European Review for Medical and Pharmacological Sciences 2018; 22: 55-61 Expression of three in endometrioid adenocarcinoma and their significance in clinical nursing, diagnosis and treatment

X.-Y. YIN1, X. YANG2, Y. CUI1, S.-M. ZHANG3

1Department of Gynecology, Affiliated Hongqi Hospital of Mudanjiang Medical University, Mudan Jiang City, Heilongjiang Province, China 2Department of Orthopedic, Affiliated Hongqi Hospital of Mudanjiang Medical University, Mudan Jiang City, Heilongjiang Province, China 3Nursing Department, Affiliated Hongqi Hospital of Mudanjiang Medical University, Mudan Jiang City, Heilongjiang Province, China

Abstract. – OBJECTIVE: This study aimed to Introduction investigate the role of NEDD9 (neural precursor cell expressed developmentally down-regulated As one of the three major malignant tumors 9), BCAR1/P130CAS (BCAR1/P130 Crk-associat- affecting the health in women, endometrial can- ed substrate) and paxillin in predicting the prog- cer is commonly manifested as adenocarcinoma1. nosis of endometrioid adenocarcinoma (EA), so as to guild the nursing of EA. The most common type of adenocarcinoma is PATIENTS AND METHODS: A total of 65 pa- endometrioid carcinoma, which can easily infil- tients who visited Affiliated Hongqi Hospital of trate into the deep muscular layer and spread to Mudanjiang Medical University between June the lymph nodes, thus leading to high mortality 2015 and June 2017 were enrolled. They under- of the patients2-4. Therefore, it is important to went gynecological surgery and had their EA investigate the factors and molecular markers confirmed by pathology, and they were assigned associated with the metastasis and recurrence of to the EA group. All EA tissues were sampled endometrial carcinoma, so as to elucidate their and archived in paraffin blocks. In addition, 40 specimens of atypical endometrial hyperplasia mechanisms. Such study will also be conducive (EAH) (the EAH group) and 40 specimens of nor- to personalized treatment and prognosis of endo- mal proliferative endometria with benign uter- metrial carcinoma. ine fibroids (the EN group) were selected as NEDD9 is a skeleton containing many controls. The protein levels of NEDD9, BCAR1/ protein interacting domains5,6. It plays a “router” P130CAS, and paxillin in each group were then function in mediating cell signaling7,8 and is detected by immunohistochemical staining. crucial in the regulation of cell transfer, chemo- RESULTS: The expression of the three pro- teins in the EA group and EAH group was sig- taxis, , , and differentiation. nificantly higher than that in the EN group, BCAR1/P130CAS has a kinase activ- 9 10 and their expression was significantly correlat- ity at the binding sites of V-Src and V-Crk , ed with the clinical stage, histological grade which plays an important role in the metastasis and lymph node metastasis of EA. In addition, and proliferation of tumor11-13. Widely recog- the expression of NEDD9, BCAR1/P130CAS, and nized as a type of structural proteins14,15 paxillin paxillin in the EA group was positively correlat- is a tyrosine phosphorylated protein found in ed with each other. cells transfected with viral oncogenes and plays CONCLUSIONS: BCAR1/P130CAS and paxillin interact with NEDD9 to participate in the growth a very important role in local adhesion. In ad- and migration of EA cells. Therefore, their pro- dition, the abnormal expression of paxillin is teins can be used as biomarkers for the diagno- related to the occurrence and development of tu- sis, treatment, and prognosis of EA. mor16-18. However, few studies have investigated the individual role of these proteins and their in- Key Words: teractions in endometrial carcinoma. Therefore, Endometrioid adenocarcinoma, NEDD9, BCAR1/ P130CAS, Paxillin. this work will investigate the role of these three proteins in the occurrence, development, migra-

Corresponding Author: Shumin Zhang, MD; e-mail: [email protected] 55 X.-Y. Yin, X. Yang, Y. Cui, S.-M. Zhang tion, infiltration, and recurrence of endometrial of stage III. Furthermore, 25 cases had lymphatic carcinoma, so as to evaluate their potential in metastasis, whereas 40 cases had no lymphatic the diagnosis, treatment, and prognosis of endo- metastasis. In 27 cases, tumor infiltration in metrial carcinoma. uterine myometrium was < 50%, whereas tumor infiltration in uterine myometrium was > 50% in 38 cases. Patients and Methods

General Information Materials and Instruments 65 patients (29-73 years old) who visited Af- Primary antibodies (mouse anti-human poly- filiated Hongqi Hospital of Mudanjiang Medical clonal antibodies against NEDD9, BCAR1/ University between June 2015 and June 2017 P130CAS and paxillin) (Beijing Boisynthe- were enrolled. They underwent gynecological sis Biotechnology Co., Ltd., Beijing, China); surgery and had their endometrioid adenocar- Secondary antibodies (Eli VisionTM plus assay cinoma (EA) confirmed by pathology. Before kit, Polymer Enhancer); Reagent B-enzyme-la- the surgery, the patients received no chemo- beled anti-mouse/rabbit polymer (Polymerized therapy, radiotherapy and endocrine therapy. HRP-Anti Ms/RbIg G) (Beijing Boisynthesis Complete clinical data were available for each Biotechnology Co., Ltd., Beijing, China); DAB patient, and all EA tissues were sampled and (3,3’-diaminobenzidine) color kit (Beijing So- archived in paraffin blocks. In addition, 40 larbio Technology Co. Ltd., Beijing, China); specimens of endometrial atypical hyperplasia Phosphate Buffered Solution (PBS) (Shanghai (EAH) (the EAH group) and 40 specimens of CaMon Biological Technology Co. Ltd., Shang- normal proliferative endometria with benign hai, China); Hematoxylin-Eosin (Shanghai Har- uterine fibroids (the EN group) were selected ing Biotechnology Co. Ltd., Shanghai, China); as controls. Before collecting the samples, the Citrate antigen repair solution (Suzhou Jiqian written informed consent was obtained from Chemical Technology Co., Ltd., Suzhou City, all patients or their family members. This clini- Jiangsu, China); Adhesive slide (Shenzhen Kang cal trial was approved by the Ethics Committee Ruide Medical Technology Co., Ltd., Shenzhen at Affiliated Hongqi Hospital of Mudanjiang Guangzhou, China). Medical University. During hospitalization, Photographic microscope; Optical microscope all patients received basic care, environmental (Shanghai Chua Kang Optical Instrument Co., care, psychological intervention, pain therapy Ltd., Shanghai, China); Rotary slicer (Guang- and complications care according to their ac- zhou Yingyi Xin automation equipment Co., Ltd., tual conditions. In addition, the patients were Guangzhou, Guangdong, China); Slice roast ma- encouraged to perform light exercise after the chine (Hubei Taikang Medical Equipment Co., surgery, so as to promote the venous return in Ltd., Xiaogan, Hubei, China); Constant tempera- the lower extremities and to prevent deep vein ture oven; Constant temperature water bath; Au- thrombosis. tomatic drift film; Disposable blade; Incubator After collection, all specimens were fixed in (Shanghai Kaye laboratory equipment Co., Ltd., 10% neutral formalin, followed by routine dehy- Shanghai, China). dration, paraffin embedding and archiving. Sub- sequently, the paraffin blocks were cut into sec- Hematoxylin-eosin (HE) Staining tions of 4 μm in thickness and coated with 0.05% The sections were subjected to conventional polylysine. To ensure the accuracy of results, all xylene dewaxation and gradient ethanol hydra- biopsy samples were diagnosed by two experi- tion, followed by 5 min hematoxylin staining enced physicians independently. According to and rinsing with tap water. Subsequently, the the Federation International of Gynecology and sections were treated for 1-3 sec in 1% hydro- Obstetrics (FIGO), the specimens were graded chloric acid and rinsed with tap water for 2 min. and staged. Among the specimens, there were 20 In the next step, the sections were stained in 1% cases of well-differentiated (G1) adenocarcinoma, of ammonia for 10 sec, followed by eosin stain- 19 cases of moderately differentiated (G2) adeno- ing for 3 min. Finally, the sections were dehy- carcinoma and 26 cases of poorly differentiated drated by gradient alcohol, treated in xylene and (G3) adenocarcinoma. In addition, there were 16 mount in neutral gum before routine pathologic cases of Stage I, 13 cases of stage II, and 36 cases observation.

56 Expression of three proteins in EA and their significance

Immunohistochemical Staining by sented by brownish yellow or chocolate brown Streptavidin-peroxidase (SP) particles located in the cytoplasm. The positive First, the PBS buffer, citrate antigen repair staining of paxillin was represented by brownish solution, and 3% hydrogen peroxide solution were yellow or chocolate brown particles located in prepared. Each tissue sample was sliced into 5 the cytoplasm and/or cell membrane. Under the sections and 3 of them were stained with NEDD9, high-power magnification, 10 view fields were BCAR1/P130CAS, and paxillin, respectively. randomly selected from each slide. If the average Among these 3 sections, one was stained with number of positive cells in each view field was >, HE, and the other one was taken as a negative 10%, the slide was deemed positive; otherwise, control. The specific steps were as follows: the slide was deemed negative. The tissue pieces were continuously sliced into sections with a thickness of 4 μm and placed on Statistical Analysis slides, which were placed in a constant tempera- All the data were first sorted using Excel and ture oven of 72°C for 1 h. The antigens on the then analyzed using statistical software SPSS sections were repaired with the citrate buffer and 19.0. The correlation among the protein expres- rinsed three times with 0.1 M PBS (pH 7.4) for sion of NEDD9, BCAR1/P130CAS, and paxil- 3 min. 3% hydrogen peroxide solution (finished lin was evaluated by Spearman rank correlation product) was added onto the slides and incubated analysis. p < 0.05 indicates the difference was at room temperature for 15 min to block the activ- statistically significant. ity of endogenous peroxidase, followed by rinsing in 0.1M PBS. Subsequently, 5%-10% of normal goat non-immune serum was dropped onto the Results sections and incubated at room temperature for 15 min, followed by incubation with the primary Characteristics of Protein Expression antibody (the dilution ratio of NEDD9 was 1:20; The expression of NEDD9 and BCAR1/ the dilution ratio of BCAR1/P130CAS was 1:100; P130CAS was located in the cytoplasm, where- and the dilution ratio of paxillin was 1:20) in as the expression of paxillin was located in the a wet box overnight at 4°C. After rinsing with cytoplasm and/or cell membrane. The EN group 0.1M PBS, the slides were dried before a polymer showed no staining or mild staining of the three enhancer (reagent A) was added dropwise onto proteins, whereas positive staining of the proteins the slides and incubated at room temperature for was observed in EA and EAH groups. The spe- 15 min. After rinsing with 0.1M PBS, the slides cific distribution of protein expression is shown were dried and incubated with enzyme-labeled in Figure 1. anti-mouse/rabbit polymer (reagent B) at room temperature for 15 min. After rinsing with 0.1M The Expression of Three Proteins PBS, the slides were dried and incubated with The expression rate of NEDD9 protein in the DAB solution for 2-3 min, followed by he- the EA, EAH and EN groups was 72.308%, matoxylin staining for 4 min. The sections were 62.500%, and 22.500%, respectively. There subsequently treated using hydrochloric acid/al- was no significant difference in NEDD9 ex- cohol and stained blue with ammonia, followed pression between EAH and EA groups (p > by 5 min sequential treatment in 85% ethanol I, 0.05). However, the expression rate of NEDD9 95% ethanol I, 95% ethanol I, anhydrous ethanol protein in EAH and EA groups was signifi- I and anhydrous ethanol II, and 2 min treatment cantly higher than that in the EN group (p < in xylene I and xylene II, respectively. Finally, 0.05). The expression rate of BCAR1/P130CAS the slides were mounted in neutral gum. Positive in EA, EAH and EN groups was 70.769%, and negative controls were also prepared during 55.000%, and 17.500%, respectively. The ex- staining, in which PBS instead of primary anti- pression rate of BCAR1/P130CAS in EAH and body was used as the negative control. EA groups was not statistically different (p > 0.05). However, the expression rate of BCAR1/ The Criteria for Diagnosis P130CAS protein in EAH and EA groups was The HE-stained sections were observed under significantly higher than that in the EN group a microscope to determine the pathological type (p < 0.05). The expression rate of paxillin of tissue samples. In particular, the positive stain- in EA, EAH and EN groups was 69.231%, ing of NEDD9 and BCAR1/P130CAS was repre- 57.500% and 22.500%, respectively. There was

57 X.-Y. Yin, X. Yang, Y. Cui, S.-M. Zhang

Figure 1. Distribution of three proteins in tissues (A, paxillin; B, BCAR1/P130CAS; C, NEDD9; 1, endometrial adenocarcinoma; 2, endometrial atypical hyperplasia; 3, normal endometrial tissue) (IHC 200×). no significant difference in the expression rate protein in EAH and EA groups was significant- of paxillin between EAH and EA groups (p > ly higher than that in the EN group (p < 0.05). 0.05). However, the expression rate of paxillin The detailed data are shown in Table I.

Table I. Expression of three proteins in different groups.

Number Positive Group of cases Positive Negative rate (%) p

NEDD9 EA group 65 47 18 72.308 0.0822* EAH group 40 25 15 62.500 0.029# EN group 40 9 31 22.500 0.006& BCAR1/P130CAS EA group 65 46 19 70.769 0.0615* EAH group 40 22 18 55.000 0.026# EN group 40 7 33 17.500 0.003& Paxillin EA group 65 45 20 69.231 0.0713* EAH group 40 23 17 57.500 0.035# EN group 40 9 31 22.500 0.007&

Note: *means that EA was compared with EAH; #means that EAH was compared with EN; &means that EA was compared with EN.

58 Expression of three proteins in EA and their significance

The Relationship Between the

Expression of Three Proteins and p > 0.05 < 0.05 < 0.05 < 0.05 the Clinical Features of EA < 0.05 The positive expression of NEDD9, BCAR1/ P130CAS and paxillin in different clinical stages, 7 7 17 17 13 13 histological grades and myometrial invasion was 13 statistically different (p < 0.05). The expression Paxillin of NEDD9, BCAR1/P130CAS, and paxillin in samples with lymph node metastasis was signifi- Negative cantly higher than that in samples with no lymph 31 16 14 22 23 30 6 22 23 3 23 22 3 node metastasis (p < 0.05). Nevertheless, the ex- Positive pression of the three proteins was not statistically different among EA samples from different age p > 0.05 < 0.05 < 0.05 < 0.05 groups (p > 0.05). The detailed data are shown < 0.05 in Table II.

9 9 The Correlation Between the Expression 5 17 16 15 13

of Three Proteins The relationship between the expression of

NEDD9, BCAR1/P130CAS, and paxillin in EA BCAR1/P130CAS was analyzed by Spearman test. The correlation Negative analysis showed that the expression of the three 14 14 21 33 3 33 26 23 23 3 23 proteins was positively correlated with each other 23 4 (r = 0.512, p < 0.01; r = 0.453, p < 0.05; and r = Positive 0.552, p < 0.01, respectively). p > 0.05 < 0.05 < 0.05 < 0.05 < 0.05 Discussion 8 4 17 17 14 10 Previous studies19-21 have found that NEDD9 15 NEDD9 can promote and invasion. From Negative the results of this work, it can be seen that NEDD9 is involved in the process of transforma-

13 13 27 22 25 1 25 20 34 22 2 3 tion involving benign and malignant endometrial 34

tissues. Therefore, NEDD9 can be used as a Positive predictor for the invasion and metastasis of EA. It may also act as a new target for the treatment of EA.

BCAR1/P130CAS plays a crucial role in cell 35 39 38 29 36 30 27 26 25 40 adhesion and extracellular matrix22,23. Cabodi of cases et al24 found that the elevated expression of Number BCAR1/P130CAS protein could accelerate the proliferation of mammary epithelial cells and promote the development of tumor. Frolov et al ≤ 55 > 55 G1+G2 G3 I+II III Negative Positive < 1/2 25 found that in glioblastoma, BCAR1/P130CAS ≥ 1/2 is involved in the vascular endothelial receptor signaling pathway, thereby pro- moting tumor invasion and metastasis. Paxillin can bind to v-Src, v-Crk and many other tum- origenic proteins, thereby affecting the growth Category

factor signaling cascades required for cell pro- The relationship between the expression three of proteins and the clinical features endometrioid of adenocarcinoma. liferation and promoting tumor metastasis26-28. In this study, it was found that the expression

Age

FIGO grading

FIGO staging Lymphatic metastasis Myometrial invasion of BCAR1/P130CAS and paxillin in EA and II. Table

59 X.-Y. Yin, X. Yang, Y. Cui, S.-M. Zhang

EAH groups was significantly higher than that 7) Li P1, Sun T2, Yuan Q1, Pan G1, Zhang J1, Sun D. in the EN group, and the expression level was The expressions of NEDD9 and E-cadherin cor- closely related to the clinical stage, histological relate with metastasis and poor prognosis in tri- ple-negative breast cancer patients. Onco Tar- grade and lymph node metastasis of EA. If the gets Ther 2016; 19: 5751-5759. clinical stage and histological grade were higher 8) Shagisultanova E, Gaponova AV, Gabbasov R, Nico- with lymph node metastasis, the expression of las E, Golemis EA. Preclinical and clinical studies BCAR1/P130CAS and paxillin was high. There- of the NEDD9 scaffold protein in cancer and oth- fore, both BCAR1/P130CAS and paxillin can act er diseases. 2015; 567: 1-11. as new targets for EA treatment. 9) Nick AM1, Stone RL, Armaiz-Pena G, Ozpolat B, Te- kedereli I, Graybill WS, Landen CN, Villares G, Vi- vas-Mejia P, Bottsford-Miller J, Kim HS, Lee JS, Kim SM, Baggerly KA, Ram PT, Deavers MT, Coleman RL, Conclusions Lopez-Berestein G, Sood AK. Silencing of p130Cas in ovarian carcinoma: a novel mechanism for tu- BCAR1/P130CAS and paxillin can interact mor cell death. J Natl Cancer Inst 2011; 103: with NEDD9 to participate in cell growth and 1596-1612. migration. They can interact with Focal Adhe- 10) Schrecengost RS, Riggins RB, Thomas KS, Guerrero sion Kinase (FAK), v-Src and v-Crk to promote MS, Bouton AH. Breast cancer antiestrogen resis- tance-3 expression regulates breast cancer cell cell proliferation and migration. Therefore, the migration through promotion of p130Cas mem- expression of these three markers in EA can be brane localization and membrane ruffling. Can- used to predict prognosis and to guide the clinical cer Res 2007; 67: 6174-6182. care, diagnosis, and treatment of EA. 11) Li L, Qu YW, Li YP. Over-expression of miR-1271 inhibits endometrial cancer cells proliferation and induces cell apoptosis by targeting CDK1. Eur Rev Med Pharmacol Sci 2017; 21: 2816-2822. Conflict of Interest 12) Rufanova VA, Alexanian A, Wakatsuki T, Lerner A, The Authors declare that they have no conflict of interests. Sorokin A. Pyk2 mediates endothelin-1 signal- ing via p130Cas/BCAR3 cascade and regulates human glomerular mesangial and References spreading. J Cell Physiol 2009; 219: 45-56. 13) Cabodi S, Tinnirello A, Di Stefano P, Bisarò B, Am- brosino E, Castellano I, Sapino A, Arisio R, Caval- Liu W, Zhang B, Xu N, Wang MJ, Liu Q 1) . miR-326 lo F, Forni G, Glukhova M, Silengo L, Altruda F, regulates EMT and metastasis of endometrial Turco E, Tarone G, Defilippi P. p130Cas as a new cancer through targeting TWIST1. Eur Rev Med regulator of mammary epithelial cell prolifera- Pharmacol Sci 2017: 21: 3787-3793. tion, survival, and HER2-neu oncogene-depen- 2) Karadayi N, Gecer M, Kayahan S, Yamuc E, Onak NK, dent breast tumorigenesis. Cancer Res 2006; 66: Korkmaz T, Yavuzer D. Association between human 4672-4680. papillomavirus and endometrial adenocarcinoma. 14) Chen DL, Wang DS, Wu WJ, Zeng ZL, Luo HY, Qiu Med Oncol 2013; 30: 597. MZ, Ren C, Zhang DS, Wang ZQ, Wang FH, Li YH, 3) Zhang HM, Fan TT, Li W, Li XX. Expressions and Kang TB, Xu RH. Overexpression of paxillin in- significances of TTF-1 and PTEN in early endo- duced by miR-137 suppression promotes tumor metrial cancer. Eur Rev Med Pharmacol Sci 2017; progression and metastasis in . 21: 20-26. Carcinogenesis 2013; 34: 803-811. 4) Castellvi J, Garcia A, Ruiz-Marcellan C, Hernán- 15) van Zyp Jv, Conway WC, Craig DH, van Zyp Nv, Tham- dez-Losa J, Peg V, Salcedo M, Gil-Moreno A, Ramon ilselvan V, Basson MD. Extracellular pressure stim- y Cajal S. in endometrial carcinoma: ulates tumor cell adhesion in vitro by paxillin acti- phosphorylated 4E-binding protein-1 expression vation. Cancer Biol Ther 2006; 5: 1169-1178. in endometrial cancer correlates with aggres- 16) Chiu HY, Sun KH, Chen SY, Wang HH, Lee MY, Tsou sive tumors and prognosis. Hum Pathol 2009; 40: YC, Jwo SC, Sun GH, Tang SJ. Autocrine CCL2 1418-1426. promotes cell migration and invasion via PKC 5) Zhang S, Wu L, Liu Q, Chen K, Zhang X. Study on activation and tyrosine of pax- effect of the expression of siRNA in gastric can- illin in bladder cancer cells. Cytokine 2012; 59: cer bearing nude mice transplanted tumor NEDD9 423-432. gene. Pak J Pharm Sci 2014; 27: 1651-1656. 17) Huang SM, Hsu PC, Chen MY, Li WS, More SV, Lu 6) Singh MK, Izumchenko E, Klein-Szanto AJ, Egleston KT, Wang YC. The novel indole compound SK228 BL, Wolfson M, Golemis EA. Enhanced genetic in- induces apoptosis and FAK/Paxillin disruption in stability and dasatinib sensitivity in mammary tu- tumor cell lines and inhibits growth of tumor graft mor cells lacking NEDD9. Cancer Res 2010; 70: in the nude mouse. Int J Cancer 2012; 131: 722- 8907-8916. 732.

60 Expression of three proteins in EA and their significance

18) Zhang LL, Mu GG, Ding QS, Li YX, Shi YB, Dai JF, Yu lesser-known CAS protein family members. Gene HG. Phosphatase and Tensin Homolog (PTEN) 2015; 570: 25-35. represses colon cancer progression through in- 24) Cabodi S, Tinnirello A, Di Stefano P, Bisarò B, Am- hibiting paxillin transcription via PI3K/AKT/NF-κB brosino E, Castellano I, Sapino A, Arisio R, Caval- pathway. J Biol Chem 2015; 290: 15018-15029. lo F, Forni G, Glukhova M, Silengo L, Altruda F, 19) Kondo S, Iwata S, Yamada T, Inoue Y, Ichihara H, Turco E, Tarone G, Defilippi P. p130Cas as a new Kichikawa Y, Katayose T, Souta-Kuribara A, Yamaza- regulator of mammary epithelial cell prolifera- ki H, Hosono O, Kawasaki H, Tanaka H, Hayashi tion, survival, and HER2-neu oncogene-depen- Y, Sakamoto M, Kamiya K, Dang NH, Morimoto C. dent breast tumorigenesis. Cancer Res 2006; Impact of the signaling adaptor protein 66: 4672-4680. NEDD9 on prognosis and metastatic behavior of 25) Frolov A, Evans IM, Li N, Sidlauskas K, Paliashvi- human lung cancer. Clin Cancer Res 2012; 18: li K, Lockwood N, Barrett A, Brandner S, Zachary 6326-6338. IC, Frankel P. Imatinib and Nilotinib increase glio- 20) Chen D, Jiao XL, Liu ZK, Zhang MS, Niu M. Knock- blastoma cell invasion via Abl-independent stim- down of PLA2G2A sensitizes gastric cancer cells ulation of p130Cas and FAK signalling. Sci Rep to 5-FU in vitro. Eur Rev Med Pharmacol Sci 2016; 6: 27378. 2013; 17: 1703-1708. 26) Huang SM, Hsu PC, Chen MY, Li WS, More SV, Lu 21) Bradshaw LN, Zhong J, Bradbury P, Mahmassani M, KT, Wang YC. The novel indole compound SK228 Smith JL, Ammit AJ, O’Neill GM. Estradiol stabilizes induces apoptosis and FAK/Paxillin disruption in the 105-kDa phospho-form of the adhesion dock- tumor cell lines and inhibits growth of tumor graft ing protein NEDD9 and suppresses NEDD9-de- in the nude mouse. Int J of Cancer 2012; 131: 722- pendent cell spreading in breast cancer cells. 732. Biochim Biophys Acta 2011; 1813: 340-345. 27) Yang WJ, Zhong J, Yu JG, Zhao F,Xiang Y. The 22) Wallez Y, Riedl S J, Pasquale E B. Association of structure and functions of paxillin and its roles in the Breast Cancer Antiestrogen Resistance Pro- neovascularization. Eur Rev Med Pharmacol Sci tein 1 (BCAR1) and BCAR3 scaffolding proteins in 2017; 21: 1768-1773. cell signaling and antiestrogen resistance. J Biol 28) Li YJ, Zhu HX, Zhang D, Li HC, Ma P, Huang LY. Chem 2014; 289: 10431-10444. Novel endogenous negative modulators of plate- 23) Deneka A, Korobeynikov V, Golemis E A. Embryonal let function as potential anti-thrombotic targets. Fyn-associated substrate (EFS) and CASS4: The Eur Rev Med Pharmacol Sci 2017; 21: 3146-3158.

61