摘要
目的 研究SERPINA5异常表达对食管鳞癌细胞化疗敏感性的影响及分子机制。方法 采用Western blot检测SERPINA5食管黏膜上皮细胞系SHEE及食管鳞癌细胞系KYSE150、TE-1、ECA109的表达水平。选取KYSE150/TE-1细胞系转染过表达空载慢病毒(oe-NC)、过表达SERPINA5慢病毒(oe-SERPINA5),采用CCK-8检测不同浓度(0、20、40、60、80μmol/L)顺铂(Cisplatin, CDDP)处理KYSE150/TE-1细胞24、48 h的半数抑制浓度(IC_(50))。采用Edu和平板克隆实验检测过表达SERPINA5后KYSE150/TE-1细胞在顺铂作用下的增殖情况,流式细胞术实验检测细胞的凋亡比率。15只雌性裸鼠被随机分为oe-NC组、oe-NC+CDDP组和oe-SERPINA5+CDDP组,通过皮下接种转染oe-NC/oe-SERPINA5慢病毒的KYSE150细胞株构建裸鼠移植瘤模型,予以oe-NC组腹腔注射生理盐水,oe-NC+CDDP组和oe-SERPINA5+CDDP组腹腔注射等体积顺铂溶液(1次/3 d,共7次),并测量各组裸鼠瘤体体积及质量,进一步通过Ki67、TUNEL染色检测移植瘤组织增殖及凋亡水平。Western blot检测PI3K/AKT信号通路蛋白表达水平。结果 SERPINA5在食管鳞癌细胞中呈低表达。与oe-NC组比较,oe-SERPINA5组IC_(50)值明显降低(P<0.05);与oe-NC+CDDP组比较,oe-SERPINA5+CDDP组细胞增殖水平降低、凋亡比例增加,差异有统计学意义(P<0.05)。动物实验结果表明,SERPINA5过表达显著抑制顺铂处理下的瘤体生长,降低肿瘤细胞增殖能力,并促进其凋亡,差异有统计学意义(P<0.05)。与oe-NC+CDDP组比较,oe-SERPINA5+CDDP组可抑制PI3K和AKT蛋白磷酸化,差异有统计学意义(P<0.05)。结论 SERPINA5过表达可降低食管鳞癌细胞增殖能力和诱导细胞凋亡,提高食管鳞癌细胞的化疗敏感性,其机制与抑制PI3K/AKT信号通路相关。
Objective To study the effect and molecular mechanism of SERPINA5 abnormal expression on chemotherapy sensitivity of esophageal squamous cell carcinoma(ESCC)cells.Methods Western blot was used to detect the expression levels of SERPINA5 esophageal mucosal epithelial cell line SHEE and ESCC cell lines KYSE150,TE-1and ECA109.Select KYSE150/TE-1 cell lines transfected with overexpression empty vector(oe-NC)and overexpression SERPINA5 vector(oe-SERPINA5),and using CCK-8 to detect the half maximal inhibitory concentration(IC 50)of cisplatin(CDDP)at different concentrations(0,20,40,60,80μmol/L)treated KYSE150/TE-1 cells for 24 and 48 hours.Edu and plate cloning experiments were used to detect the proliferation of KYSE150/TE-1 cells overexpressing SERPINA5 under cisplatin treatment,and flow cytometry was used to detect the apoptosis rate.15 female nude mice were randomly divided into oe-NC group,oe-NC+CDDP group and oe-SERPINA5+CDDP group.A nude mouse transplant tumor model was constructed by subcutaneous inoculation of KYSE150 cell line transfected with oe-NC/oe-SERPINA5 lentivirus.The oe-NC group was intraperitoneally injected with physiological saline,while the oe-NC+CDDP group and oe SERPINA5+CDDP group were intraperitoneally injected with cisplatin solution(once/three days,a total of seven times).The tumor volume and mass of each group of nude mice were measured,and the proliferation and apoptosis levels of the transplant tumor tissue were further detected by Ki67 and TUNEL staining.Western blot was used to detect the expression levels of PI3K/AKT signaling pathway proteins.Results SERPINA5 showed low expression in ESCC cells.Compared with the oe-NC group,the IC 50 value of the oe SERPINA5 group was significantly reduced(P<0.05).Compared with the oe-NC+CDDP group,the oe-SERPINA5+CDDP group showed a significant decrease in cell proliferation and an increase in apoptosis rate(P<0.05).Animal experiment results showed that overexpression of SERPINA5 significantly inhibited tumor growth under cisplatin treatment,reduced tumor cell proliferation ability and promoted apoptosis,with statistical significance(P<0.05).Compared with the oe-NC+CDDP group,the oe-SERPINA5+CDDP group inhibited PI3K and AKT protein phosphorylation,and the difference was statistically significant(P<0.05).Conclusion Overexpression of SERPINA5 reduces the proliferation ability of ESCC cells and induces cell apoptosis,and increases chemotherapy sensitivity of esophageal squamous cell carcinoma cells.Its mechanism is related to the inhibition of the PI3K/AKT signaling pathway.
作者
李志芳
魏瑜
苏比努尔·阿不拉克
魏行方
张莉
LI Zhifang;WEI Yu;Subinuer Abulake;WEI Xingfang;ZHANG Li(Department of Comprehensive Internal Medicine,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)
出处
《新疆医科大学学报》
2025年第5期569-578,共10页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区重点研发计划项目(2020B03003,2020B03003-3)。
