摘要
目的观察龙胆苦苷对非小细胞肺癌细胞A549株化疗敏感性的影响,并探讨可能机制。方法取对数期A549/DDP细胞,随机分为对照组(常规培养基)、龙胆苦苷组(培养基含有龙胆苦苷40μmol/L)、IGF-1组(培养基含有IGF-1100μg/L)、联合组(培养基含有龙胆苦苷40μmol/L及IGF-1100μg/L)。培养48 h进行后续实验。MTT法检测细胞增殖抑制率;流式细胞术检测细胞凋亡率;MDC染色检测细胞自噬小体;Western blot法检测细胞微管相关蛋白1轻链3融合蛋白Ⅱ(LC3Ⅱ)、p62/SQSTM1、蛋白激酶B(Akt)、p-Akt、哺乳动物雷帕霉素靶蛋白(mTOR)、p-mTOR蛋白表达量。结果与对照组比较,龙胆苦苷组增殖抑制率、凋亡率、MDC阳性率、LC3Ⅱ蛋白表达量升高,p62/SQSTM1蛋白表达量、p-AKT/AKT、p-mTOR/mTOR降低(P<0.05),IGF-1组增殖抑制率、凋亡率、MDC阳性率、LC3Ⅱ蛋白表达量降低,p62/SQSTM1蛋白表达量、p-AKT/AKT、p-mTOR/mTOR升高(P<0.05);与龙胆苦苷组比较,联合组增殖抑制率、凋亡率、MDC阳性率、LC3Ⅱ蛋白表达量降低,p62/SQSTM1蛋白表达量、p-AKT/AKT、p-mTOR/mTOR升高(P<0.05);与IGF-1组比较,联合组增殖抑制率、凋亡率、MDC阳性率、LC3Ⅱ蛋白表达量升高,p62/SQSTM1蛋白表达量、p-AKT/AKT、p-mTOR/mTOR降低(P<0.05)。结论龙胆苦苷可诱导非小细胞肺癌耐药细胞自噬并提升其化疗敏感性,抑制PI3K/Akt信号通路可能是其作用机制之一。
Objective To observe the effect of gentiopicroside on the chemotherapy sensitivity of non-small cell lung cancer cell line A549,and to explore the possible mechanism.Methods A549/DDP cells in log phase were randomly divided into control group(conventional medium),gentiopicroside group(medium containing 40μmol/L gentiopicroside),IGF-1 group(medium containing IGF-1100μg/L),combined group(medium containing 40μmol/L gentiopicroside and IGF-1100μg/L).Cultivate for 48 h for follow-up experiments.The MTT method was used to detect the inhibition rate of cell proliferation.Flow cytometry was used to detect the rate of apoptosis.MDC staining was used to detect autophagosomes.Western blot was used to detect cell microtubule-associated protein 1 light chain 3 fusion protein II(LC3 II),p62/SQSTM1,protein kinase B(Akt),p-Akt,mammalian target of rapamycin(mTOR),p-mTOR protein expression.Results Compared with the control group,the proliferation inhibition rate,apoptosis rate,MDC positive rate,LC3Ⅱprotein expression were increased,p62/SQSTM1 protein expression,p-AKT/AKT,p-mTOR/mTOR were decreased in the gentiopicroside group(P<0.05),the proliferation inhibition rate,apoptosis rate,MDC positive rate,LC3Ⅱprotein expression were decreased,and p62/SQSTM1 protein expression,p-AKT/AKT,and p-mTOR/mTOR were increased in the IGF-1 group(P<0.05).Compared with the gentiopicroside group,the proliferation inhibition rate,apoptosis rate,MDC positive rate,LC3Ⅱprotein expression were decreased,and p62/SQSTM1 protein expression,p-AKT/AKT,p-mTOR/mTOR were increased in the combined group(P<0.05).Compared with the IGF-1 group,the proliferation inhibition rate,apoptosis rate,MDC positive rate,LC3Ⅱprotein expression were increased,and p62/SQSTM1 protein expression,p-AKT/AKT,p-mTOR/mTOR were decreased in the combined group(P<0.05).Conclusion Gentiopicroside can induce autophagy in non-small cell lung cancer resistant cells and enhance their chemotherapy sensitivity,and inhibition of PI3K/Akt signaling pathway may be one of its mechanisms.
作者
刘瀚文
王旸
杨峥
LIU Hanwen;WANG Yang;YANG Zheng(Nanyang Central Hospital,Nanyang,473000)
出处
《实用癌症杂志》
2023年第9期1398-1402,共5页
The Practical Journal of Cancer
关键词
龙胆苦苷
磷脂酰肌醇3激酶
蛋白激酶B
自噬
非小细胞肺癌
化疗敏感性
Gentiopicroside
Phosphatidylinositol 3-kinase
Protein kinase B
Autophagy
Non-small cell lung cancer
Chemotherapy sensitivity
