QIAN Liyong,YANG Zhiqiang,WEN Yuanyuan. Over expression of SIAH1 induced apoptosis of breast cancer cells by up-regulating the level of Hrk[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2018, 48(1): 20-25,33.
Abstract：Objective: To investigate SIAH1 and Hrk expression in breast cancer, analyze the relationship between expression and clinicopathological features, and explore the expression SIAH1 promoting apoptosis of breast cancer by up-regulating Hrk expression. Methods: In this study, the Hrk and SIAH1 protein expression were detected in 86 cases of breast invasive ductal carcinoma and 32 cases in normal tissue adjacent to breast cancer by immunohistochemical SP method. The expression of Hrk and SIAH1 were examined in MCF-10A (breast normal epithelial cell line) and MCF-7 (breast cancer cell line). In MCF-7, it was transfected with pcDNA3-myc-SIAH1 (SIAH1 overexpression plasmid) only, and co-transfected with Hrk siRNA and pcDNA3-myc-SIAH1 by liposome-mediated method, respectively. Then cell apoptosis was detected with Annexin V-FITC/PI reagent kit and Flow cytometry assay. Results: Immunohistochemical analysis showed that the expression of SIAH1 and Hrk were lower in breast invasive ductal carcinoma tissues in comparison with normal tissues adjacent to carcinoma. The expression level of SIAH1 and Hrk correlated with higher grade (χ2=8.403, 8.514, P<0.01) and lower TNM stage (χ2=6.731, 6.729, P<0.05) significantly in breast cancer, and both the expression was positively correlated (r=0.515, P<0.01). After overexpressing SIAH1, the mRNA and protein expression of SIAH1 and Hrk (P<0.05), and the number of breast cancer cells apoptosis increased (29.02%±3.31%, P<0.05). Co-transfection pcDNA3-myc-SIAH1 and Hrk siRNA (4.40%±0.61%), compared with separate transfection pcDNA3-myc-SIAH1 (29.02%±3.31%), breast cancer cells apoptosis rate dropped significantly (F=187.369, P<0.05). Conclusion: Overexpression of SIAH1 can induce apoptosis of breast cancer cells by up-regulating the level of Hrk.
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