摘要
目的研究罗格列酮对人肝癌HepG2细胞增殖与凋亡的影响,探讨p38丝裂原活化蛋白激酶(p38MAPK)通路相关蛋白在其中发挥的作用。方法 MTT法检测罗格列酮对人肝癌HepG2细胞的增殖抑制率,流式细胞术检测细胞凋亡率,透射电子显微镜观察细胞凋亡形态变化,Western blot检测p38MAPK通路相关蛋白表达变化。结果罗格列酮可抑制HepG2细胞的增殖,诱导细胞凋亡(P<0.05);电子显微镜下可观察到典型的HepG2细胞凋亡表现。Western blot结果显示罗格列酮可激活p38MAPK通路,上调HepG2细胞中磷酸化p53及Bax蛋白的表达,下调Bcl-2蛋白的表达(P<0.05);而细胞外信号调节激酶ERK1/2的磷酸化程度没有明显改变。p38MAPK通路抑制剂SB203580可显著降低罗格列酮诱导的HepG2细胞的凋亡率;并且SB203580可部分逆转由罗格列酮引发的磷酸化p53、Bax及Bcl-2蛋白的表达变化。结论罗格列酮可通过激活p38MAPK通路诱导人肝癌HepG2细胞凋亡,其机制可能与p38MAPK通路参与磷酸化p53、Bax及Bcl-2蛋白的调控有关。
Objective To investigate the effects of Rosiglitazone(ROZ) on the proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 and to explore the role of p38MAPK signaling pathway in these processes. Methods HepG2 cell proliferation was determined by MTT method, and the apoptosis rates were measured by flow cytometry(FCM). The morphology of cell apoptosis was investigated by transmission electron microscopy(TEM). Western blot was performed to measure the protein expression related to p38MAPK signaling pathway. Result Rosiglitazone inhibited HepG2 cell proliferation in a concentration- and time-dependent manner(P<0.05), and the apoptosis rates were increased significantly(P<0.05). The typical morphological changes of apoptosis were chromatin condensation and aggregation at the periphery of nucleons and nuclear fragmentation. Rosiglitazone activated p-p38 MAPK pathway, upregulated the expression of p-p53 and Bax, and down-regulated Bcl-2 expression in HepG2 cells(P<0.05), while with no effect on ERK1/2 phosphorylation. The rosiglitazone-induced apoptosis of HepG2 cells and the changes of p-p53, Bcl-2 and Bax proteins were partly blocked by p38MAPK inhibitor SB203580. Conclusion Rosiglitazone could activate p38MAPK pathway by inhibiting HepG2 cell proliferation and inducing its apoptosis. The mechanism may be related with p38MAPK signaling pathway regulating the expression of p-p53, Bcl-2 and Bax proteins.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2014年第11期1181-1185,共5页
Cancer Research on Prevention and Treatment
基金
河北省高校强势特色学科项目[2005(52)]
河北省卫生厅资助课题(06142)
