siRNA干扰S100A4对人胰腺癌细胞增殖和凋亡及其对吉西他滨敏感性影响的研究

Effect of silence S100A4 by RNA interference on proliferation,apoptosis,chemosensitivity of gemcitabine of the pancreatic cancer cell lines

目的:观察利用小干扰RNA(siRNA)手段下调人胰腺癌BxPC-3、AsPC-1细胞中S100A4表达后对细胞凋亡、增殖、吉西他滨化疗敏感性的影响。方法:设计合成针对S100A4特异的siRNA转染人胰腺癌BxPC-3、AsPC-1细胞,以RT-PCR检测转染前后S100A4蛋白表达变化;转染前后BxPC-3细胞生长曲线检测BxPC-3、AsPC-1细胞增殖能力;TUNEL法检测转染前后细胞凋亡变化;MTT检测不同浓度吉西他滨对胰腺癌细胞半数抑制浓度(IC50)。结果:RT-PCR结果显示BxPC-3、AsPC-1细胞转染S100A4siRNA 48h后,S100A4mRNA表达明显下调;S100A4siRNA转染后BxPC-3、AsPC-1细胞增殖下降;TUNEL结果显示转染后凋亡明显增加;MTT结果显示,BxPC-3细胞对照组吉西他滨IC50为(9.95±0.34)mmol/L;阴性转染组吉西他滨IC50为(9.641±0.434)mmol/L;干扰组吉西他滨IC50为(1.182±0.175)μmol/L;AsPC-1细胞对照组吉西他滨IC50(64.15±3.41)mmol/L;阴性转染组吉西他滨IC50为(65.19±4.4)mmol/L;干扰组吉西他滨IC50为(1.962±0.113)mmol/L,P<0.05。结论:S100A4沉默后抑制胰腺癌细胞增殖、促进凋亡并提高吉西他滨化疗敏感性,提示S100A4可能是治疗胰腺癌的有效靶点。

OBJECTIVE： To observe the effect on the proliferation, apoptosis and chemosensitivity of gemcitabine after slience S100A4 by RNA interference. METHODS： the S100A4-speeified small interference RNA was designed and synthesized. The expression of S100A4 was detected by RT-PCR 24 h after the S100A4 siRNA transfected into BxPC-3 and AsPC-1 cells. The growth curve of BxPC-3 and AsPC-1 cells were recorded to demonstrate the proliferation. The ap- optosis of BxPC-3 and AsPC-1 cells was tested by flow cytometric analysis. The gemcitabine of IC50 was measured by MTT. RESULTS： S100A4 expression was suppressed significantly after S100A4 siRNA transfected into BxPC-3 and As- PC-1 cells. The abilities of proliferation were inhibited significantly after S100A4 siRNA transfected. TUNEL assay showed apoptosis increased after transfection. Before and after slience S100A4 by siRNA, IC50 of gemcitabine in control was （9.95±0.34） mmol/L,Negative Control was （9. 641±0. 434） mmol/L, Interference was （1. 182±0. 175）μmol/L, respectively; in AsPC-1, control was （64.15±3.41） mmol/L,Negative Control was （65. 19±4.4） mmol/L,Interference was （1. 962±0. 113） retool/L, respectively. CONCLUSIONS： S100A4 slience can inhibite cell proliferation and enhence the apoptosis as well as increase the chemosensitivity of gemcitabine. S100A4 could serve as a potential target in the filed of pancreatic cancer therapy.