摘要
目的:确定转录活化因子3(STAT3)抑制剂HO-3867与顺铂单独或联合使用对子宫颈癌HeLa细胞生物学行为的影响,并探索其作用机制。方法:培养子宫颈癌HeLa细胞,采用噻唑蓝(MTT)法检测不同浓度的HO-3867(0~10μmol/L)和顺铂(0~20μg/ml)对HeLa细胞增殖的影响,根据结果确定实验浓度,将子宫颈癌HeLa细胞随机分为4组:对照组(无顺铂及HO-3867)、HO-3867组(3.69μmol/L)、顺铂组(4.9μg/ml)、联合组(2μmol/L HO-3867+2.5μg/ml顺铂)。采用MTT法检测细胞的增殖抑制率;平板克隆实验检测细胞的克隆形成能力;流式细胞术检测细胞的凋亡情况;Transwell小室检测细胞的侵袭能力;划痕实验检测细胞的迁移能力;Western Blot方法检测细胞JAK1/STAT3信号通路、凋亡、侵袭与迁移相关蛋白的表达情况。结果:与对照组比较,顺铂组、HO-3867组、联合组的增殖抑制率和总凋亡率显著增加(P<0.01),克隆形成能力、穿膜细胞数及迁移能力显著降低(P<0.01);且联合组较顺铂组和HO-3867组效果更加显著(P<0.01)。与对照组比较,HO-3867组、顺铂组、联合组细胞中Bcl-2、MMP-2、p-JAK1、p-STAT3蛋白的表达明显降低(P<0.01),而Bax、E-cadherin蛋白的表达明显升高(P<0.01);并且联合组对上述蛋白的调控作用更加明显(P<0.01)。结论:HO-3867与顺铂联合使用通过抑制JAK1/STAT3信号通路关键蛋白的磷酸化,进而抑制子宫颈癌HeLa细胞的增殖、克隆形成、迁移及侵袭能力,促进其凋亡;且HO-3867的使用可减少顺铂的使用剂量,间接减少了顺铂的毒副反应和耐药。
Objective: To determine the effect of STAT3 inhibitor HO-3867 and cisplatin alone or in combination on biological behavior of cervical cancer Hela cells and explore its mechanism. Methods: Cervical cancer He La cells were cultured. The effects of different concentrations of HO-3867( 0-10 μmol/L) and cisplatin( 0-20 μg/ml) on the proliferation of Hela cells were detected by MTT assay. The experimental concentration was determined according to the results. Hela cells were randomly divided into four groups: control group( no cisplatin and HO-3867),HO-3867 group( containing 3. 69 μmol/L HO-3867),cisplatin group( containing 4. 9 μg/ml cisplatin) and combination group( containing 2 μmol/L HO-3867 + 2. 5 μg/ml cisplatin). Cell proliferation inhibition rate was measured by MTT assay;cell clonogenic ability was measured by plate cloning assay;cell apoptosis was measured by flow cytometry assay;and cell invasion was measured by Transwell chamber;Western Blot was used to detect the expression of proteins related to apoptosis,invasion and migration. Results: Compared with the control group,the proliferation inhibition rate and total apoptosis rate of cisplatin group,HO-3867 group and combined group were significantly increased( P < 0. 01),and the colony forming ability,the number of transmembrane cells and migration ability were significantly decreased( P < 0. 01). The combined group was more effective than cisplatin group and HO-3867 group( P < 0. 01). Compared with the control group,the expression of Bcl-2,MMP-2,p-JAK1,p-STAT3 in the cells of HO-3867 group,cisplatin group and combined group decreased significantly( P < 0. 01),while the expression of Bax and E-cadherin increased significantly( P < 0. 01). And the regulation effect of the combined group on the above proteins was more obvious( P < 0. 01). Conclusions: HO-3867 combined with cisplatin can inhibit the proliferation,cloning,migration and invasion of Hela cells by inhibiting the phosphorylation of JAK1/STAT3 signaling pathway and promote their apoptosis. HO-
作者
任倩梅
郭倩
詹平
毛熙光
REN Qianmei;GUO Qian;ZHAN Ping(Department of Obstetrics and Gynecology,Affiliated Hospital of Southwest Medical University,Luzhou Sichuan 646000,China)
出处
《实用妇产科杂志》
CAS
CSCD
北大核心
2022年第1期73-78,共6页
Journal of Practical Obstetrics and Gynecology
