Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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On Sep 2018




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On Sep 2018




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"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
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Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
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Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
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Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
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Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2022 | Month : October | Volume : 16 | Issue : 10 | Page : EC01 - EC06 Full Version

Evaluation of CD9 Expression of Tumour Cells and Stromal Immune Cells in Breast Carcinoma by Immunohistochemistry


Published: October 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/57978.17002
RV Sowjanya Lakshmi, ML Harendra Kumar, k Mohan Kumar

1. Postgraduate Student, Department of Pathology, Sri Devaraj Urs Medical College, Kolar, Karnataka, India. 2. Professor, Department of Pathology, Sri Devaraj Urs Medical College, Kolar, Karnataka, India. 3. Professor, Department of General Surgery, Sri Devaraj Urs Medical College, Kolar, Karnataka, India.

Correspondence Address :
Dr. ML Harendra Kumar,
Professor, Department of Pathology, Sri Devaraj Urs, Tamaka, Kolar, Karnataka, India.
E-mail: harendrakmr929@gmail.com

Abstract

Introduction: Cluster of Differentiation 9 (CD9), known as Motility Related Protein (MRP-1) regulates cell adhesion, motility, migration and proliferation. Many studies have stated conflicting results on prognostic significance of invasive breast carcinomas with CD9 expression that had performed on tumour tissues.

Aim: To assess the inter-relationship of CD9 expression of tumour cells and stromal immune cells in breast carcinoma with clinicopathological parameters which include age, tumour size, grade, histological type, lymph nodes, tumour staging and molecular classification.

Materials and Methods: An observational prospective (July 2020 to June 2021) and retrospective (October 2019 to June 2020) study was done in 71 cases of resected primary invasive breast carcinoma over a period of one and half year at Sri Devaraj Urs Medical College, Karnataka, India. Immunohistochemistry (IHC) staining was done using CD9 antibody. Tumour cells (T-CD9) expression was evaluated by Immunoreactivity scoring (IS= Intensity score×Extent of staining). The stromal cells (S-CD9) expression was evaluated by percentage (%) of stromal area occupied by CD9 stained immune cells. Chi-square test was used as test of significance for qualitative data. The p-value of <0.05 was considered as statistically significant.

Results: Out of 71 cases, T-CD9 expression was noticed in 40 (56.34%) cases and IS <4 considered as negative was observed in 31 (43.66%) cases. However there was no association with age, tumour size, grade and molecular markers for the expression of both T-CD9 and S-CD9. Human Epidermal growth factor receptor 2 neu (HER2neu) negative was associated with T-CD9 expression (p-value=0.05). Hence, CD9 can be used as prognostic marker for Her2neu negative cases.

Conclusion: The CD9 expression was not significantly associated with tumour cells (T-CD9) and stromal cells (S-CD9) in breast carcinoma cases. However, it was significantly associated with Her2neu negative tumour cells. T-CD9 showed more positivity in Luminal A followed by triple negative, whereas S-CD9 showed more positivity in Luminal B. CD9 did not show association with any parameters except Her2neu negative.

Keywords

Invasive breast cancer, Molecular classification, Motility related protein-1, Nottingham prognostic index scoring, Stromal cells

Breast cancer has become the most commonly diagnosed cancer worldwide surpassing lung cancer and is one of the leading sources of cancer associated deaths (1). During 2020, there were 2.3 million women diagnosed with breast cancer with global mortality rate of 685,000 (1). There were 1,78361 new cases and 90,408 deaths in India during 2020 (2). The incidence rate of breast cancer in Kolar region was 6.41% in 2010 (3).

The CD9, otherwise known as Motility Related Protein (MRP-1) which is a member of tetraspanin family associated in various process that includes cell adhesion, cell motility, migration and proliferation as a result of interaction with integrins and other tetraspanins (CD8 and CD151), growth factor receptors and signalling molecules (4). It is a cell membrane glycoprotein and it has four functional regions which are N and C terminal cytoplasmic domains, a small intracellular loop and 2 extracellular loops (4),(5). CD9 is extensively detected in various types of cancer cells and also in normal tissues (6). Previous studies have showed that decreased expression of CD9 would be associated with poor prognosis in breast carcinoma and various other malignancies such as oesophageal carcinoma, oral squamous cell carcinoma and gastrointestinal stromal tumour (7),(8),(9),(10). Various studies have stated conflicting results on prognostic significance of invasive breast carcinomas with CD9 expression that was performed on tumour tissues (6),(7). The purpose of present study was to evaluate CD9 expression in tumour cells as well as stromal immune cells by performing IHC in invasive breast carcinoma cases and to evaluate the inter-relationship of tumour cells-CD9 and stromal cells-CD9 expression with clinicopathological parameters and with molecular classification.

Material and Methods

An observational prospective (July 2020 to June 2021) and retrospective (October 2019 to June 2020) study done in duration of one and half year at Sri Devaraj Urs Medical College at Tamaka, Kolar, India, was conducted.

Primary invasive breast carcinoma that had undergone surgical resection were collected from the archives of pathology in R.L. Jalappa Hospital and Research Centre attached to Sri Devaraj Urs Medical College, Tamaka, Kolar, Karnataka after obtaining the approval from Ethics Committee (DMC/KLR/IEC/694/2020-21).

Inclusion criteria: In the present study, mainly infiltrating ductal carcinoma cases and other histological types such as papillary carcinoma, invasive lobular carcinoma, mucinous carcinoma, medullary carcinoma, metaplatsic carcinoma, phyllodes tumour and mixed tumours were included in the study.

Exclusion criteria: All the cases of breast carcinoma patients who received neoadjuvant chemotherapy or any other malignancies and small biopsies were excluded from this study.

Sample size calculation: Sample size of 71 was based on CD9 expression in study by Baek J et al., (42.5%) with 95% confidence intervals with absolute error of 10% (11). Sample size formula: N=Zα2 P (1-P)/d2, P=expected proportion i.e. 42.5%, Zα=1.96 (95% confidence interval), D=absolute error (10%).

Study Procedure

All the Haematoxylin and Eosin (H&E) slides were rescreened by two authors to confirm the histological type, tumour grade, presence and absence of nodal metastasis and lymphovascular invasion. IHC slides of Estrogen Receptor (ER), Progesterone Receptor (PR), Her2 and Ki-67 were also screened. All the breast carcinoma cases were classified according to World Health Organisation (WHO) classification (12). Tumour grading was done as per Modified Scarff- Bloom Richardson Histologic Grading (13).

Patients survival status was determined using Nottingham Prognostic Index (NPI) and was calculated using standard formula NPI = (0.2xS)+G+N

where S= Tumour Size in cm, G= Grade of tumour (grade 3=3, grade 2=2, grade 1=1), N=Number of lymph nodes involved (>4=3, 4-1=2, 0=1). All cases were categorised into three different prognostic groups: Good=2.00-3.40, moderate=3.41-5.40, poor=>5.40 (14). IHC was done using principle of peroxidase-antiperoxidase method using tonsil as positive control. ER, PR was interpreted as per the Allred score and Her2neu were scored as per 2018 American Society of Clinical Oncology (ASCO) guidelines (15),(16). Ki-67 was scored using the standard guidelines (17). To evaluate the CD9 expression IHC was performed by using CD9 antibody. In both tumour cells (T-CD9) as well as stromal cells (S-CD9), CD9 expression was elucidated. To elucidate the T-CD9 expression IS was used (Table/Fig 1). This IS scoring is produced by taking intensity and extent of staining into consideration and is as follows (11),(18):

On the basis of mean IS, cases were divided into positive and negative and the scoring is as follows: IS : <4: Negative, ≥4: Positive. The stromal immune cells (S-CD9) expression was evaluated by: Percentage (%) of stromal area occupied by CD9 stained immune cells. S-CD9 was assessed same as Tumour Infiltrating Lymphocytes (TILs) (19). Molecular classification of all cases was done and T-CD9 and S-CD9 expression were analysed.

Molecular classification: By performing IHC on the tumour tissues of breast, cases were classified into five subtypes depending on the hormonal status as stated in 2011 St. Gallen consensus is as follows (Table/Fig 2) (20).

To assess the prognosis NPI scoring system and other clinicopathological parameters which include age, tumour size, grade, lymph node, pathological Tumour, Node and Metastases (TNM) staging, hormonal markers and molecular classification were used and evaluated the association of both T-CD9 and S-CD9 expression.

Statistical Analysis

Data was entered into Microsoft excel data sheet and was analysed using Statistical Package for the Social Sciences (SPSS) 22.0 version software (IBM SPSS Statistics, Somers NY, USA). Categorical data was represented in the form of frequencies and proportions. Chisquare test or Fisher’s exact test (for 2×2 tables only) was used as test of significance for qualitative data. The p-value (probability that the result is true) of ≤0.05 was considered as statistically significant.

Results

Out of 71 cases of invasive breast carcinomas, CD9 expression was elucidated by IHC in T-CD9 and S-CD9 and these were interpreted and assessed separately. Of all cases, the intensity for T-CD9 expression was differed for every case that is 28 (39.4%) cases showed strong positivity (Table/Fig 3), moderate positivity was observed in 24 (33.8%) cases, 10 (14.1%) cases showed weak positivity and 9 (12.7%) cases were showed negative. Also this CD9 expression was noticed in normal breast ductal epithelial cells which were expressed as weak positivity. CD9 was expressed in both tumour cells and stromal immune cells (Table/Fig 4).

In 71 cases, CD9 expression in tumour cells with IS >4 which considered positive was noticed in 40 (56.34%) cases and IS <4 which considered as negative was observed in 31 (43.66%) cases. Among 71 cases, 57 were of Infiltrating Ductal Carcinoma (IDC) type and 14 cases were of other histological types. Out of these 57 IDC cases, T-CD9 was expressed in 31 (54.4%) cases and 26 (45.6%) cases did not show any expression and among 14 other histological types, 9 (64.29%) cases showed T-CD9 expression and 5 (35.71%) cases did not show T-CD9 expression. Other histological types which showed T-CD9 expression were mixed carcinoma (IDC+ILC) (Table/Fig 5), mucinous carcinoma (Table/Fig 6), papillary carcinoma (Table/Fig 7), invasive lobular carcinoma (Table/Fig 8), medullary and metaplastic carcinoma (Table/Fig 9). T-CD9 was not expressed in phyllodes tumour (Table/Fig 10). Whereas, S-CD9 expression was seen in 43 (60.56%) cases and S-CD9 expression was not observed in 28 (39.44%) cases (Table/Fig 11). However there was no association observed with clinicopathological data such as age, tumour size, tumour grade and molecular markers of ER, PR, and Ki67 for the expression of both T-CD9 and S-CD9 (Table/Fig 12),(Table/Fig 13). But Her2neu molecular marker was associated with T-CD9 expression with p-value=0.05 (Table/Fig 13).

Discussion

Breast carcinoma is one of the most common cancers worldwide (1). Because of the extension of the tumour tissue and heterogeneous nature of the breast tumour metastases, the morbidity is increasing in breast carcinoma cases. The major concern about the breast carcinoma is its metastatic spread. To invade into mesenchymal stromal cells, breast cancer cells depend on CD9.

Many studies demonstrated that decreased expression of CD9 correlated with a poor prognosis in breast carcinomas (7). In a study done by Rappa G et al., it was reported that the invasion of breast cancer cells into mesenchymal stromal cells was reduced with CD9 lacking breast cancer cells and this study suggested that by inhibiting CD9, metastases and invasion can be reduced (21).

In this study, CD9 was expressed in both tumour cells and stromal immune cells and their expression patterns have different association with clinicopathological features of patients with invasive breast carcinoma. In the present study, among 71 cases T-CD9 positive expression was observed in 40 cases. Out of these 31 cases were of invasive ductal carcinoma and nine cases were other histological types which include two mixed carcinoma cases (IDC+ILC), two mucinous carcinomas, two papillary carcinoma, one each of ILC, medullary and metaplastic carcinoma types. Remaining 31 cases showed negative T-CD9 expression. Among these 26 were IDC type and five cases were of other histological type which include three cases of mixed carcinomas (ILC +Mucinous, IDC+ Mucinous, IDC+ILC) and two phyllodes tumours.

In the study, done by Jamil F et al., it was noted that strong CD9 expression was seen in all normal and benign epithelial cells and 40% of ductal carcinoma in situ and IBCs. The rest of the tumour cases exhibited weak CD9 expression. There was no significant association of CD9 expression with pathological parameters and also with molecular markers such as tumour grading, lymph node metastases and ER, PR respectively. They had stated that CD9 marker could not serve as a significant prognostic marker in invasive breast carcinoma cases (22). Present study showed similar results as the expression of T-CD9 marker was not significantly associated with clinicopathological parameters and with molecular markers except Her2neu negative marker.

The expression of CD9 showed weak intensity in benign ductal epithelial cells and normal breast tissues in this study. In this study the prognostic significance of CD9 expression was also compared with NPI scoring system and did not show any significant association (14).

The study done by Adams S et al., showed the expression of stromal immune cells in invasive breast carcinoma cases had better prognostic significance especially with molecular classification that includes mainly triple negative breast carcinoma cases (23). In contrary to their study, present study did not show any significant association for stromal immune cells (S-CD9) with any clinicopathological parameters and molecular classification.

In the study done by Khomo S et al., CD9 was expressed more in chemotherapy resistant cases, disseminated spread and also in recurrent cases of lung carcinoma. This could be used for targeted therapy. When coming to metastases of breast carcinomas especially to the bone with comparison of primary breast carcinoma cases and visceral metastases, the expression of CD9 was considered significantly more in bone metastases (24).

In the study done by Kischel P et al., both prostate carcinomas and osteotropic breast carcinoma cases CD9 was excessively expressed (25).

Limitation(s)

Present study was a unicentric study done in a smaller sample size. The sample size of other histological types was also less. IHC for CD9 was not performed on metastatic lymph nodes.

Conclusion

The immunostaining for CD9 expression was not significantly associated with Tumour cells (T-CD9) and Stromal immune cells (S-CD9) in breast carcinoma cases. However, it was significantly associated with Her2neu negative for T-CD9. Hence, CD9 can be used as prognostic marker for Her2neu negative cases. CD9 expression for tumour cells showed more positivity in Luminal A followed by triple negative whereas S-CD9 showed more positivity in Luminal B. Assessment of CD9 expression depends on many variables like genetic makeup and variability, tumour microenvironment and molecular pathogenesis in our geographical population. However, furthermore follow-up studies, genetic studies and compartment specific studies of CD9 expression in invasive breast carcinomas are needed for prognostic significance as there are no studies reported in India.

References

1.
Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. Ca Cancer J Clin. 2021;71:209-49. Doi: 10.3322/caac.21660. [crossref] [PubMed]
2.
Cancer today [Internet]. Gco.iarc.fr. 2022 [cited 27 July 2022]. Available from: https://gco.iarc.fr/today/ fact-sheets- populations.
3.
Kalyani R, Das S, Bindra Singh M, Kumar HML. Cancer profile in Kolar: A ten years study. Indian J Cancer. 2010;2:160-65. [crossref] [PubMed]
4.
Lazo PA. Functional implications of tetraspaninproteins in cancer biology. Cancer Sci. 2007;98:1666-77. [crossref] [PubMed]
5.
Stipp CS, Kolesnikova TV, Hemler ME. Functional domains in tetraspanin proteins. Trends Biochem Sci. 2003;28:106-12. [crossref] [PubMed]
6.
Huang C, Taki T, Adachi M, Yagita M, Sawada S, Takabayashi A, et al. MRP-1/ CD9 and KAI1/CD82 expression in normal and various cancer tissues. Int J Oncol. 1997;11:1045-51. [crossref] [PubMed]
7.
Miyake M, Nakano K, Itoi S, Koh T, Taki T. Motility-related protein-1 (MRP-1/ CD9) reduction as a factor of poor prognosis in breast cancer. Cancer Res. 1996;56;1244-49.
8.
Uchida S, Shimada Y, Watanabe G, Li ZG, Hong T, Miyake M, et al. Motilityrelated protein (MRP-1/CD9) and KAI1/CD82 expression inversely correlate with lymph node metastasis in oesophageal squamous cell carcinoma. Br J Cancer. 1999;79:1168-73. [crossref] [PubMed]
9.
Buim ME, Lourenço SV, Carvalho KC, Cardim R, Pereira C, Carvalho AL, et al. Downregulation of CD9 protein expression is associated with aggressive behavior of oral squamous cell carcinoma. Oral Oncol. 2010;46:166-71. [crossref] [PubMed]
10.
Soyuer S, Soyuer I, Unal D, Ucar K, Yildiz OG, Orhan O, et al. Prognostic significance of CD9 expression in locally advanced gastric cancer treated with surgery and adjuvant chemo radiotherapy. Pathol Res Pract. 2010;206:607-10. [crossref] [PubMed]
11.
Baek J, Jang N, Choi J, Kim J, Bae Y. CD9 expression in tumor cells is associated with poor prognosis in patients with invasive lobular carcinoma. J Breast Cancer. 2019;22(1):77. [crossref] [PubMed]
12.
Lakhani SR. International Agency for Research on Cancer. WHO classification of breast tumours: WHO classification of tumours, volume 2. 2nd ed. Who Classification of Tumours Editorial Board, editor. IARC; 2019.
13.
Meyer JS, Alvarez C, Milikowski C, Olson N, Russo I, Russo J, et al. Breast carcinoma malignancy grading by Bloom–Richardson system vs proliferation index: Reproducibility of grade and advantages of proliferation index. Mod Pathol. 2005;18:1067-78. [crossref] [PubMed]
14.
Zhen H, Yang L, Li L, Yu J, Zhao L, Li Y, et al. Correlation analysis between molecular subtypes and Nottingham prognostic index in breast cancer. Onco Target. 2017;8:74096-05. [crossref] [PubMed]
15.
Allred DC, Harvey JM, Berardo M. Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol.1998;11:155-68.
16.
Kim MC, Kang SH, Choi JE, Bae YK. Impact of the updated Guidelines on Human Epidermal Growth Factor 2 (HER 2) Testing in breast cancer. J Breast Cancer. 2020;23:484-97. [crossref] [PubMed]
17.
Nahed A, Shaimaa M. Ki-67 as a prognostic marker according to breast cancer molecular subtype. Cancer Biology and Medicine. 2016;13(4):496. [crossref] [PubMed]
18.
Kwon H, Choi J, Kang S, Son Y, Bae Y. Prognostic significance of CD9 expression differs between tumour cells and stromal immune cells, and depends on the molecular subtype of the invasive breast carcinoma. Histopathology. 2017;70(7):1155-65. [crossref] [PubMed]
19.
Salgado R, Denkert C, Demaria S, Sirtaine N, Klauschen F, Pruneri G, et al. The evaluation of tumour-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015;26(2):259-71. [crossref] [PubMed]
20.
Mittal A, Mani N. Molecular classification of breast cancer. Indian J Pathol Oncol. 2021;8(2):241-47. [crossref]
21.
Rappa G, Green TM, Karbanova J, Corbeil D, Lorico A. Tetraspanin CD9 determines invasiveness and tumorigenicity of human breast cancer cells. Oncotarget. 2015;6:7970-91. [crossref] [PubMed]
22.
Jamil F, Peston D, Shousha S. CD9 immunohistochemical staining of breast carcinoma:unlikely to provide useful prognostic information for routine use. Histopathology. 2001;39:572-77. [crossref] [PubMed]
23.
Adams S, Gray RJ, Demaria S, Goldstein L, Perez EA, Shulman LN, et al. Prognostic value of tumour-infiltrating lymphocytes in triple-negative breast cancers from two phase III randomized adjuvant breast cancer trials: ECOG 2197 and ECOG 1199. J Clin Oncol. 2014;32:2959-66. [crossref] [PubMed]
24.
Kohmo S, Kijima T, Otani Y, Masahide M, Minami T, Takahashi R, et al. Cell surface tetraspanin CD9 mediates chemo resistance in small cell lung cancer. Cancer Res. 2010;70:8025-35. [crossref] [PubMed]
25.
Kischel P, Bellahcene A, Deux B, Lamour V, Dobson R, De Pauw E, et al. Overexpression of CD9 in human breast cancer cells promotes the development of bone metastases. Anticancer Res. 2012;32:5211-20.

DOI and Others

DOI: 10.7860/JCDR/2022/57978.17002

Date of Submission: May 23, 2022
Date of Peer Review: Jun 28, 2022
Date of Acceptance: Aug 16, 2022
Date of Publishing: Oct 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? Yes
• For any images presented appropriate consent has been obtained from the subjects. Yes

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Jun 01, 2022
• Manual Googling: Aug 09, 2022
• iThenticate Software: Aug 13, 2022 (11%)

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