西罗莫司通过MAPK信号通路抑制人宫颈癌细胞增殖的实验研究

Experimental study of rapamycin inhibiting proliferation of human cervical cancer cells through MAPK signaling pathway

目的探讨雷帕霉素抑制人宫颈癌细胞Hela增殖的机制。方法 0、10、30、100nmol/L的西罗莫司作用Hela细胞24、48、72h,MTT法检测细胞活力,流式细胞术检测细胞凋亡及细胞周期,Western blot检测细胞中B淋巴细胞瘤-2基因(Bcl-2)蛋白、Bcl-2相关X蛋白(Bax)、细胞周期蛋白D1(Cyclin D1),p21,丝裂原活化蛋白激酶(MAPK)信号通路相关蛋白的表达。结果与0nmol/L比较,10、30、100nmol/L西罗莫司能显著降低细胞活力(P<0.01),提高细胞凋亡率(P<0.01),使细胞周期阻滞于G1期(P<0.01),下调Cyclin D1,Bcl-2,磷酸化细胞外信号调节激酶(p-ERK)1/2,磷酸化c-Jun氨基末端激酶(p-JNK)及p-p38MAPK表达(P<0.01),上调Bax及p21表达(P<0.01)。与西罗莫司组比较,西罗莫司+SB203580组、西罗莫司+SP600125、西罗莫司+PD98059组Cyclin D1,Bcl-2,p-ERK1/2、p-p38 MAPK及p-JNK表达下调(P<0.01),Bax及p21表达上调(P<0.01)。结论西罗莫司通过阻断MAPK信号通路抑制Hela细胞增殖。

Objective To investigate the mechanism of rapamycin inhibiting the proliferation of human cervical cancer cells-Hela.Methods Hela cells were cultured with 0,10,30,100 nmol/L dose of rapamycin for 24,48,72 h.Cell viability was detected by MTT assay.Apoptosis and cell cycle were detected by flow cytometry.The expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),CyclinD1,p21 and MAPK signaling pathway related proteins were measured by Western blot.Results Compared with 0 nmol/L,10,30,100 nmol/L rapamycin could reduce cell viability(P<0.01),increase apoptosis rate(P<0.01),and arrest cell cycle in G1 phase(P<0.01),down-regulation the expression of Cyclin D1,Bcl-2,phosphorylation of extracellular signal-regulated kinase(p-ERK)1/2,p-JNK and p-p38 MAPK(P<0.01),up-regulation the expression of Bax and p21(P<0.01)significantly.Compared with the rapamycin group,the expression of CyclinD1,Bcl-2,p-ERK1/2,p-JNK and p-p38 MAPK was down-regulated(P<0.01),the expression of Bax and p21 was up-regulated in the rapamycin+SB203580,rapamycin+SP600125 and rapamycin+PD98059 group.Conclusion Rapamycin inhibits the proliferation of Hela cells by blocking the MAPK signaling pathway.