隐丹参酮联合顺铂抗非小细胞肺癌NCI-H1975细胞JAK2/STAT3机制研究

摘要

Objective The aim of this study was to investigate the synergistic anti-tumor effect of cryptotanshinone and cis-platin in non-small cell lung cancer NCI-H1975 cells and its possible molecular mechanism. Methods NCI-H1975 cells were treated with control,cryptotanshinone,cisplatin or combination of cryptotanshinone and cisplatin groups(referred to as the combination group). The inhibitory rate of cell proliferation was determined in NCI-H1975 cells by CCK-8 assay;Flow cytometry was used to determine the rates of survival and cell apoptosis;The expression of apoptotic protein,anti-apoptotic protein,JAK2/STAT3 protein and its phosphorylation levels were detected in NCI-H1975 cells by Western blot. The localization and transcription activity of STAT3 cells were determined by laser confocal microscopy/luciferase assays. Results 1. The survival rates in cryptotanshinone,cisplatin and combination groups at each time-point were lower in NCI-H1975 cells than that in the control group(P<0. 05);The proliferation of NCI-H1975 cells in the combination group were inhibited when compared to the control,cryptotanshinone,or cisplatin groups(P<0. 05);2. After treatments for 24 h,the expression levels of Bcl-2 and Survivin protein were decreased(P<0. 01),the activity of Caspase3 and Caspase9 protein was increased(P<0. 01),and the expression levels of p-STAT3 and p-JAK2 protein were de-creased in NCI-H1975 cells(P<0. 01);3. After NCI-H1975 cells treated with the combination of cryptotanshinone and cisplatin, the STAT3 in the nucleus was decreased,and STAT3 activity was also decreased in the nucleus. Conclusion Cryptotanshinone com-bined with cisplatin can exert a synergistic antitumor effect on non-small cell lung cancer NCI-H1975 cells,and its mechanism is related to the inhibition of JAK2 and STAT3 phosphorylation signal pathways.%目的 探讨隐丹参酮与顺铂联合作用于非小细胞肺癌NCI-H1975细胞具有协同抗肿瘤效应及其可能的分子机制.方法 实验分为对照组、隐丹参酮组、顺铂组及隐丹参酮和顺铂联合用药组(以下简称联合用药组),CCK-8法测定细胞增殖抑制率,流式细胞术测定细胞存活率及凋亡,免疫印迹技术检测用药后凋亡蛋白、抗凋亡分子蛋白表达水平、JAK2/STAT3蛋白及其磷酸化表达水平,激光共聚焦/荧光素酶检测STAT3细胞定位和转录活性.结果 (1)在各时间点,单药组及联合用药组存活率均低于对照组(P<0. 05);与对照组及单药组相比,联合用药组在各时间点均抑制NCI-H1975细胞增殖活性(P<0. 05);(2)联合用药组作用于NCI-H1975细胞24 h后,抗凋亡蛋白Bcl-2、Survivin表达水平下降(P<0. 01),Caspase3和Caspase9活性提高(P<0. 01),p-STAT3及p-JAK2表达水平下降(P<0. 01);(3)联合用药组作用于NCI-H1975细胞后,激光共聚焦检测提示细胞核内STAT3明显减少,核内STAT3活性降低.结论 隐丹参酮联合顺铂作用于非小细胞肺癌NCI-H1975细胞能发挥协同抗肿瘤作用,其机制与抑制信号通路JAK2/STAT3磷酸化表达水平有关.