학술논문
山柰酚通过Met/PI3K/Akt/mTOR通路诱导非小细胞肺癌NCI-H1650细胞发生自噬进而影响其增殖 / Kaempferol promotes cell autophagy and affects cell proliferation of non-small cell lung cancer H1650 cells via regulating Met/PI3K/Akt/mTOR pathway
Document Type
Academic Journal
Author
朱志明; 周绮纯; 王娟娟; 邓芷茵; 唐青; 吴万垠; 王晰; 万信良; 莫瀚丹; 王苏美; ZHU Zhiming; ZHOU Qichun; WANG Juanjuan; DENG Zhiyin; TANG Qing; WU Wanyin; WANG Xi; WAN Xinliang; MO Handan; WANG Sumei
Source
中国肿瘤生物治疗杂志 / Chinese Journal of Cancer Biotherapy. 30(11):965-972
Subject
Language
Chinese
ISSN
1007-385X
Abstract
目的:探讨山柰酚诱导人非小细胞肺癌(NSCLC)NCI-H1650细胞发生自噬及其机制.方法:常规培养NCI-H1650细胞,用不同浓度山柰酚处理细胞,用CCK-8法、MTT法以及EdU法检测山柰酚对NCI-H1650细胞增殖能力的影响,用自噬双标记腺病毒mCherry-EGFR-LC3感染实验检测山柰酚对NCI-H1650细胞发生自噬的影响,用WB法检测山柰酚处理后NCI-H1650细胞中自噬相关蛋白及Met/PI3K/Akt/mTOR信号通路相关蛋白的表达,用qPCR法检测山柰酚处理后NCI-H1650细胞中Met mRNA的表达.采用荧光素酶标记A549-luc细胞建立裸鼠移植瘤模型后用山柰酚进行处理,用活体动物成像技术观察移植瘤生长情况,用WB法检测移植瘤组织中自噬相关蛋白以及Met/PI3K/Akt/mTOR信号通路相关蛋白的表达.结果:山柰酚能显著抑制NCI-H1650细胞的增殖能力(P<0.05),山柰酚处理后NCI-H1650细胞内的自噬小体数量明显增加(P<0.05)、自噬标志蛋白LC3B和beclin1表达均明显上调(均P<0.05)、P62表达明显下调(P<0.05),山柰酚可明显抑制NCI-H1650细胞中Met mRNA和蛋白的表达(均P<0.05)、抑制p-PI3K p85、PI3K p85、p-Akt和p-mTOR蛋白表达(均P<0.05).山柰酚抑制A549细胞裸鼠移植瘤的生长(P<0.05)和影响其体内自噬、Met/PI3K/Akt/mTOR通路相关蛋白的表达(均P<0.05).结论:山柰酚通过影响Met/PI3K/Akt/mTOR通路诱导NSCLC NCI-H1650细胞发生自噬,进而抑制其增殖能力.
Objective:To explore the specific effect and mechanism of kaempferol on inducing autophagy in non-small cell lung cancer(NSCLC)NCI-H1650 cells.Methods:Human NSCLC cell line NCI-H1650 was cultured and treated with kaempferol at different concentrations.The effects of kaempferol on NSCLC cell viability and proliferation were observed by CCK-8,MTT and EdU methods.LC3 adenovirus infection experiment was performed to investigate the effect of kaempferol on NSCLC cell autophagy.Western blot was used to detect the expression of key proteins of cell autophagy and relevant molecules of Met/PI3K/Akt/mTOR signaling pathway.Meanwhile,qPCR was used to detect the mRNA expression of Met in NCI-H1650 cells after kaempferol treatment.The transplanted tumor model of nude mice was established by using luciferase labeled A549-luc cells.The tumor growth was observed by in vivo animal imaging,and the expression of autophagy related key proteins and molecules of Met/PI3K/Akt/mTOR signaling pathway in the xenograft tissues were detected by Western blot.Results:Kaempferol significantly inhibited the proliferation of NCI-H1650 cells(P<0.05).After kaempferol treatment,the number of autophagosomes in NCI-H1650 cells was significantly increased(P<0.05),the expressions of autophagy related key proteins,LC3B and beclin1,were significantly increased(all P<0.05),and the expression of P62 was significantly decreased(P<0.05).Kaempferol significantly inhibited the mRNA and protein expression of Met,and inhibited the protein expression of p-PI3K p85,PI3K p85,p-Akt and p-mTOR(all P<0.05).Kaempferol inhibited the growth of transplanted tumor(P<0.05)and affected autophagy and the expression of Met/PI3K/Akt/mTOR pathway-related proteins in nude mouse tumor models(all P<0.05).Conclusion:Kaempferol induces autophagy in NSCLC NCI-H1650 cells by affecting the Met/PI3K/Akt/mTOR pathway,which in turn inhibites their proliferative capacity.
Objective:To explore the specific effect and mechanism of kaempferol on inducing autophagy in non-small cell lung cancer(NSCLC)NCI-H1650 cells.Methods:Human NSCLC cell line NCI-H1650 was cultured and treated with kaempferol at different concentrations.The effects of kaempferol on NSCLC cell viability and proliferation were observed by CCK-8,MTT and EdU methods.LC3 adenovirus infection experiment was performed to investigate the effect of kaempferol on NSCLC cell autophagy.Western blot was used to detect the expression of key proteins of cell autophagy and relevant molecules of Met/PI3K/Akt/mTOR signaling pathway.Meanwhile,qPCR was used to detect the mRNA expression of Met in NCI-H1650 cells after kaempferol treatment.The transplanted tumor model of nude mice was established by using luciferase labeled A549-luc cells.The tumor growth was observed by in vivo animal imaging,and the expression of autophagy related key proteins and molecules of Met/PI3K/Akt/mTOR signaling pathway in the xenograft tissues were detected by Western blot.Results:Kaempferol significantly inhibited the proliferation of NCI-H1650 cells(P<0.05).After kaempferol treatment,the number of autophagosomes in NCI-H1650 cells was significantly increased(P<0.05),the expressions of autophagy related key proteins,LC3B and beclin1,were significantly increased(all P<0.05),and the expression of P62 was significantly decreased(P<0.05).Kaempferol significantly inhibited the mRNA and protein expression of Met,and inhibited the protein expression of p-PI3K p85,PI3K p85,p-Akt and p-mTOR(all P<0.05).Kaempferol inhibited the growth of transplanted tumor(P<0.05)and affected autophagy and the expression of Met/PI3K/Akt/mTOR pathway-related proteins in nude mouse tumor models(all P<0.05).Conclusion:Kaempferol induces autophagy in NSCLC NCI-H1650 cells by affecting the Met/PI3K/Akt/mTOR pathway,which in turn inhibites their proliferative capacity.