核糖体S6蛋白激酶4变异体基因表达对乳腺癌细胞增殖的影响

目的 探讨核糖体S6蛋白激酶4变异体(RSK4m)对乳腺癌细胞株MDA-MB-231增殖的影响。 方法 将RSK4m的过表达载体稳定转染至乳腺癌MDA-MB-231细胞株中,分别设一个阴性对照组(mock组)及过表达RSK4m1组(OE1组)、过表达RSK4m2组(OE2组)、过表达RSK4m3组(OE3组),采用实时荧光定量PCR(qRT- PCR)和Western blotting检测RSK4m的mRNA和蛋白的表达, CCK-8试剂盒检测转染后MDA-MB-231细胞的生长增殖, 免疫共沉淀试验(CO-IP)验证热休克蛋白与RSK4变异体的相互作用。 结果 成功转染RSK4m慢病毒表达载体至乳腺癌MDA-MB-231细胞,其mRNA和蛋白水平均过表达(P均<0.05)。OE1组、OE2组、OE3组在24、48、72、96 h的细胞抑制率(%)与mock组相比,48、72、96 h的差异具有统计学意义(P<0.05),CO-IP结果证实热休克蛋白与RSK4m存在相互作用关系。 结论 RSK4m可抑制乳腺癌细胞株MDA-MB-231的增殖,并可能与RSK4w的抑制增殖作用存在着不同的调控机制。

Effect of expression of ribosomal protein S6 Kinase 4 variants on the proliferation of breast cancer cell MDA-MB-231

Objective To investigate the effect of expression of ribosomal protein S6 kinase 4 variants(RSK4m)on the proliferation of breast cancer cell line MDA-MB-231. Methods Lentiviral vector of RSK4 variants were constructed and transfected on breast cancer cell line MDA-MB-231. A negative control group(mock group)and overexpression RSK4m1 group(OE1 group), overexpression RSK4m2 group(OE2 group), overexpression RSK4m3 group(OE3 group)were set respectively. The expressions of three RSK4 variants were detected with qRT- PCR and Western blot. The proliferation of transfected cells was assayed by Cell Counting Kit-8. Finally, the interaction of RSK4m with heat shock proteins observed in preliminary experiments was verified with CO-IP. Results The mRNA and protein expressions of RSK4 variants were significantly up-regulated in transfected cells(all P<0.05). Compared with the mock group, the OE1, OE2 and OE3 groups had significant differences in the inhibition rate(%)at 48, 72, and 96 hour(P<0.05). CO-IP results confirmed the existence of interaction between heat shock protein and RSK4m. Conclusion The expression of RSK4m can inhibit the proliferation of MDA-MB-231. Its regulatory mechanism may differ from that of RSK4w.