摘要
目的探讨LNCRNA 00593调控非小细胞肺癌(NSCLC)顺铂耐药的作用机制。方法实时荧光定量PCR(quantitative real⁃time PCR,RT⁃qPCR)检测LNCRNA 00593在癌旁正常组织(normal)、肺腺癌(LUAD)和肺鳞癌(LUSC)、HEB细胞、A549细胞、H1299细胞、PC9细胞、H460细胞和calu3细胞中的表达量;CDPP(0μM、5μM、10μM)作用H1299细胞,RT⁃qPCR检测LNCRNA 00593在顺铂治疗细胞中的表达量;10μM CDPP处理H1299细胞0、12、24、36、48 h后,RT⁃qPCR检测细胞中LNCRNA 00593的表达量。CCK⁃8及流式细胞仪分析过表达及下调LNCRNA 00593对H1299细胞顺铂耐药的影响。RT⁃qPCR及Western blot检测过表达及下调LNCRNA 00593后,H1299细胞内p53的表达量,RNA⁃pull down验证LNCRNA 00593和p53的结合。p53通路抑制剂PFT⁃α作用细胞后,检测LNCRNA 00593对顺铂诱导的H1299细胞活力和凋亡的影响。结果LNCRNA 00593在NSCLC组织的表达水平明显低于癌旁正常组织(P<0.05)。LNCRNA 00593在NSCLCA549、H1299、PC9、H460、calu3细胞中的表达水平明显低于HEB细胞(P<0.01)。分别用0μM、5μM、10μM CDPP处理H1299细胞后,LNCRNA 00593表达量随CDPP作用浓度依赖性升高(P<0.01)。用10μM CDPP处理H1299细胞12、24、36、48 h后,LNCRNA 00593表达量随CDPP作用时间不断增加(P<0.01)。CDPP作用细胞后,下调LNCRNA 00593使H1299细胞活力明显上升,凋亡率明显下降,过表达LNCRNA 00593使H1299细胞活力下降,凋亡率明显上升。过表达LNCRNA 00593激活了p53通路,LNCRNA 00593能够与p53结合。PFT⁃α作用细胞后,下调了LNCRNA 00593对H1299细胞顺铂耐药的调控作用。结论过表达LNCRNA 00593通过p53通路调控NSCLC细胞的顺铂耐药。
Objective To investigate the mechanism of LNCRNA 00593 regulating cisplatin resistancein non⁃small cell lung cancer. Methods Quantitative real⁃time PCR (RT⁃qPCR) was used to detect theexpression of LNCRNA 00593 in normal paracancer tissues (Normal), lung adenocarcinoma (LUAD) and lungsquamous cell carcinoma (LUSC), HEB cell, A549 cell, H1299 cell, PC9 cell, H460 cell and calu3 cell.H1299 cells were treated with different concentrations of CDPP (0 μM, 5 μM, 10 μM), the expression ofLNCRNA 00593 in cisplatin⁃treated cells was detected by RT⁃qPCR. After treating H1299 cells with CDPP for0 h, 12 h, 24 h, 36 h, 48 h, the expression of LNCRNA 00593 in the H1299 cells was detected by RT⁃qPCR.The effects of overexpression and down⁃regulation of LNCRNA 00593 on cisplatin resistance of H1299 cells wereanalyzed by CCK⁃8 and flow cytometry. The expression of p53 in H1299 cells was detected by RT⁃qPCR andWestern blot after overexpression and down⁃regulation of LNCRNA 00593, and the binding of LNCRNA 00593 and p53 was verified by RNA⁃pull down. The effects of LNCRNA 00593 on the viability and apoptosis of H1299cells induced by cisplatin were detected after PFT⁃α treatment. Results The expression level of LNCRNA00593 in non⁃small cell lung cancer tissues was significantly lower than that in adjacent normal tissues (P<0.05). The expression levels of LNCRNA 00593 in A549, H1299, PC9, H460 and calu3 cells weresignificantly lower than that of HEB cells (P<0.01). After treating H1299 cells with 0 μM, 5 μM, and 10 μMCDPP respectively, the expression of LNCRNA 00593 increased in a concentration⁃dependent manner (P <0.01). After treating H1299 cells with 10 μM CDPP for 12 h, 24 h, 36 h, 48 h, the expression of LNCRNA00593 increased in a time⁃dependent manner (P<0.01). After CDPP treatment, down⁃regulation of LNCRNA00593 significantly increased H1299 cell activity and decreased apoptosis rate, while overexpression ofLNCRNA 00593 significantly decreased H1299 cell activity and increased apoptosis r
作者
李峥
赵猛
葛鹏
张爱敏
任丽
Li Zheng;Zhao Meng;Ge Peng;Zhang Aimin;Ren Li(Department of Clinical Laboratory,Tumor Hospital,Tianjin Medical University,Tianjin 300060,China)
出处
《中华肺部疾病杂志（电子版）》
CAS
2020年第6期724-730,共7页
Chinese Journal of Lung Diseases(Electronic Edition)
基金
国家自然科学基金青年科学基金项目(81602026)
天津医科大学肿瘤医院科研项目(Y1806)。
