Zhang Jian, Guo Bianqin, Zhang Yan, Cao Ju, Chen Tingmei
Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Saudi Med J. 2010 Apr;31(4):374-81.
To explore the roles of annexin II in breast cancer progression, and to study the effect of annexin II on breast cancer cell proliferation and invasion.
This study was conducted in the Key Laboratory of Diagnostic Medicine Designated by the Ministry of Education, Chongqing Medical University, Chongqing, China from December 2006 to January 2009. First, we employed Western blot and reverse transcriptase polymerase chain reaction to detect the expression of annexin II and S100A10 in a panel of well-characterized human breast cancer cell lines, and investigated the localization of annexin II and S100A10 by use of immunofluorescence. We then silenced the expression of annexin II in MDA-MB-435s, which was found to over express annexin II, using the chemically-synthetic annexin II small interfering RNA (siRNA) duplexes (including 3 groups: blank MDA-MB-435s cells, cells transfected with negative control siRNA, and cells transfected with annexin II-siRNA). Finally, the cell proliferation, invasion, and plasmin generation were assayed, and the cellular levels of S100A10 and c-Myc were also detected. All the tests were repeated 3 times.
Annexin II and S100A10 were over expressed in invasive human breast cancer cell lines. The siRNA targeting annexin II of MDA-MB-435s cells did not only decrease annexin II messenger RNA and protein levels, but also down-regulated the levels of S100A10, and c-Myc. The treated cells were remarkably blocked in the G0/G1 phase, and cells in the S/G2+M phase decreased. Additionally, the treatment with siRNA resulted in reduction of plasmin generation as well as a loss of the invasive capacity of breast cancer cells.
Annexin II might be a key contributor to breast cancer proliferation and invasion.
探讨膜联蛋白II在乳腺癌进展中的作用,并研究膜联蛋白II对乳腺癌细胞增殖和侵袭的影响。
本研究于2006年12月至2009年1月在中国重庆医科大学教育部诊断医学重点实验室进行。首先,我们采用蛋白质印迹法和逆转录聚合酶链反应检测一系列特征明确的人乳腺癌细胞系中膜联蛋白II和S100A10的表达,并通过免疫荧光法研究膜联蛋白II和S100A10的定位。然后,我们使用化学合成的膜联蛋白II小干扰RNA(siRNA)双链体(包括3组:空白MDA-MB-435s细胞、转染阴性对照siRNA的细胞和转染膜联蛋白II-siRNA的细胞)沉默MDA-MB-435s中膜联蛋白II的表达,该细胞系中膜联蛋白II呈过表达。最后,检测细胞增殖、侵袭和纤溶酶生成情况,并检测S100A10和c-Myc的细胞水平。所有检测均重复3次。
膜联蛋白II和S100A10在侵袭性人乳腺癌细胞系中过表达。靶向MDA-MB-435s细胞中膜联蛋白II的siRNA不仅降低了膜联蛋白II信使核糖核酸和蛋白质水平,还下调了S100A10和c-Myc的水平。处理后的细胞在G0/G1期明显受阻,S/G2+M期的细胞减少。此外,siRNA处理导致纤溶酶生成减少以及乳腺癌细胞侵袭能力丧失。
膜联蛋白II可能是乳腺癌增殖和侵袭的关键因素。