双脱甲氧基姜黄素通过调节HIF-1α介导衰老诱导非小细胞肺癌细胞凋亡并抑制细胞增殖  

Bisdemethoxycurcumin induces apoptosis and inhibits proliferation of non⁃small cell lung cancer cells by regulating HIF⁃1α⁃mediated senescence

作　　者：刘华英 夏雪 张孟维 罗琴琴 LIU Huaying;XIA Xue;ZHANG Mengwei;LUO Qinqin(Department of Traditional Chinese Medicine,Wuhan Hospital of Traditional Chinese and Western Medicine,Wuhan NO.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市中西医结合医院,武汉市第一医院中医部,武汉430022

出　　处：《中国癌症防治杂志》2023年第3期291-297,共7页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：湖北省自然科学基金项目(2020CFB359)。

摘　　要：目的探讨双脱甲氧基姜黄素(bisdemethoxycurcumin,BDMC)对非小细胞肺癌细胞增殖和凋亡的调控作用及分子机制。方法非小细胞肺癌细胞株NCI-H1975和NCI-H520分别用磷酸盐缓冲液(PBS)和20μmol/L的BDMC处理后,采用CCK-8法检测细胞活力,羧基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescein diacetate succinimide ester,CFSE)法检测细胞增殖,吖啶橙(acridine orange,AO)染色和Annexin V-FITC/PI染色法检测细胞凋亡,细胞衰老相关β-半乳糖苷酶(senescence-associatedβ-galactosidase,SA-β-Gal)染色实验检测SA-β-Gal活性,流式细胞仪检测SA-β-Gal阳性细胞比例。然后分别以20μmol/L BDMC、20μmol/L缺氧诱导因子HIF-α抑制剂PX-478或20μmol/L BDMC+20μmol/LPX-478处理非小细胞肺癌细胞株NCI-H1975和NCI-H520,以等剂量PBS处理的细胞为对照组,采用免疫印迹法检测HIF-1α和HIF-2α蛋白的表达,SA-β-Gal染色实验检测SA-β-Gal活性。结果与PBS组相比,BDMC组NCI-H1975和NCI-H520细胞活力明显降低(均P<0.05),CFSE阳性增殖细胞比例明显减少(均P<0.01);AO阳性细胞比例明显减少(均P<0.01),Annexin V+PI-和SA-β-Gal+PI-细胞比例明显增加(均P<0.01),SA-β-Gal活性明显增加(均P<0.001),HIF-1α蛋白表达明显增加(均P<0.001)。相比于BDMC组,BDMC+PX-478组的HIF-1α蛋白表达降低(P<0.05),SA-β-Gal阳性细胞比例明显减少。结论BDMC可能通过上调HIF-1α信号促进非小细胞肺癌细胞衰老,进而抑制细胞增殖并促进凋亡。Objective To investigate the regulatory effect of dimethoxycurcumin(BDMC)on the proliferation and apoptosis in non⁃small cell lung cancer cells and its mechanism.Methods Non⁃small cell lung cancer cell lines NCI⁃H1975 and NCI⁃H520 were treated with phosphate buffered saline(PBS)and 20μmol/L BDMC.Cell viability was detected by CCK⁃8;cell proliferation was detected by carboxyfluorescein diacetate succinimide ester(CFSE);cell apoptosis was detected by acridine orange(AO)and Annexin V⁃FITC/PI staining;senescence⁃associatedβ⁃galactosidase(SA⁃β⁃Gal)staining assay was used to detect the activity of SA⁃β⁃Gal;the proportion of SA⁃β⁃Gal positive cells was detected by flow cytometer.Subsequently,NCI⁃H1975 and NCI⁃H520 cells were treated with 20μmol/L BDMC,20μmol/L hypoxia inducible factor⁃1α(HIF⁃1α)inhibitor PX⁃478,or a combination of both,respectively,and the cells treated with PBS served as the control group.The expression of HIF⁃1αand HIF⁃2αproteins was detected by Western blot,and the activity of SA⁃β⁃Gal was detected by SA⁃β⁃Gal staining assay.Results Compared with the PBS group,the cell viability of NCI⁃H1975 and NCI⁃H520 in BDMC group was significantly reduced(all P<0.05),the proportion of CFSE⁃positive proliferating cells was significantly reduced(all P<0.01),the number of AO⁃positive cells was significantly decreased(all P<0.01),the proportions of Annexin V+PI-cells and SA⁃β⁃Gal+PI-cells were significantly increased(all P<0.01),the SA⁃β⁃Gal activity was significantly increased(all P<0.001),the protein expression of HIF⁃1αwas significantly increased(all P<0.001).Compared with BDMC group,the protein expression of HIF⁃1αin BDMC+PX⁃478 group were decreased(P<0.05),as well as the proportion of SA⁃β⁃Gal⁃positive cells.Conclusions BDMC may promote senescence of non⁃small cell lung cancer cells by up⁃regulating HIF⁃1αsignaling pathway,and thus inhibit cell proliferation and promote apoptosis.

关 键 词：非小细胞肺癌 双脱甲氧基姜黄素 衰老 增殖 凋亡 缺氧诱导因子 

分 类 号：R734.2[医药卫生—肿瘤]