摘要
探讨siRNA靶向抑制Rael表达对口腔鳞状细胞癌HSC -3细胞增殖、侵袭和凋亡的影响及其机制。方法将体外培养的HSC-3细胞分为未转染组(未做任何处理)、siNC组(转染siRNA-NC)和siRacl组(转染siRNA - Rael )。 RT - PCR检测各组细胞中Rael mRNA的表达,MTT法、Transwell小室和流式细胞仪分别检测各组细胞的增殖、侵袭和凋亡情况,Western blot检测细胞中Rael蛋白、Wnt/β- catenin信号通路相关蛋白β- catenin、c - Myc及其下游凋亡相关蛋白Bel -2、Cleaved Caspase -3的表达量。结果与未转染组比较,siRacl组细胞的吸光度(0D)值明显下降,侵袭细胞数明显减少,Rael mRNA和Rael、β-catenin、c - Myc、Bel -2蛋白的表达水平均明显降低,而细胞凋亡率和Cleaved Caspase -3蛋白的表达水平明显升高(均P<O.05或0.01);而siNC组与未转染组比较无显著差异(P>0.05)。结论靶向Rael的siRNA可抑制HSC -3细胞增殖、侵袭并诱导细胞凋亡,其作用机制可能与抑制Wnt/β-catenin信号通路的活化有关。
Objective To investigate the effects of siRNA targeting Rael expression on proliferation, invasion and apoptosis of oral squamous cell carcinoma HSC - 3 cells and its mechanisms. Methods HSC - 3 cells cultured in vitro were divided into untransfected group(without any treatment),siNC group(transfected siRNA - NC) and siRacl group(transfected siRNA - Rael). The expression of Rael was detected by RT - PCR. Cell proliferation, invasion and apoptosis in each group were detected by MTT, Trans well and flow cytometry respectively. Expression of Rael protein and Wnt/β- catenin signaling pathway - related proteins β- catenin, c - Myc and downstream apoptosis - related proteins Bel - 2 and Cleaved Caspase - 3 were detected by Western blot. Results Compared with untransfected group, the OD value of cells decreased significantly, the number of invasive cells reduced markedly ,the expression levels of Rael mRNA and Rael ,β- catenin,c - Myc and Bel - 2 proteins were cut down observably, while the apoptotic rate and the expression level of Cleaved Caspase - 3 protein were increased remarkably in siRacl group(P < 0. 05 ), but there was no significant difference between siNC group and untransfected group(P > 0. 05). Conclusion siRNA targeting Rael can inhibit proliferation,invasion and induce apoptosis of HSC - 3 cells,and its mechanism may be related to inhibit the activation of Wnt/beta - catenin signaling pathway.
作者
王海业
麻颖宜
Wang Haiye;Ma Yingyi(Department of Stomatology,The First Affiliated Hospital of Zhengzhou University,ZhengZhou 450052,China)
出处
《医学新知》
CAS
2019年第3期297-301,共5页
New Medicine
