罗仙子提取物通过调控HIF1A-AS1影响结直肠癌细胞增殖、凋亡和周期分布  

Housefly maggots extracts affects the proliferation, apoptosis and cycle distribution of colorectal cancer cells by regulating HIF1A-AS1

作　　者：高海楠 金洪飞[1] 高杨[1] 杨光[1] 郝文伟[1] GAO Hai-nan;JIN Hong-fei;GAO Yang;YANG Guang;HAO Wen-wei(Department of Surgery,Tieling Central Hospital of Liaoning Province,Tieling 112000,China)

机构地区：[1]辽宁省铁岭市中心医院普外科,辽宁铁岭112000

出　　处：《中国现代普通外科进展》2021年第11期859-864,共6页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:研究罗仙子提取物对结直肠癌细胞增殖、凋亡和周期分布的影响及可能机制。方法:用中、低、高剂量的罗仙子提取物处理结肠癌细胞(SW480)24 h,实时荧光定量PCR(RT-qPCR)检测长链非编码RNA(lncRNA)低氧诱导因子1α反义RNA1(HIF1A-AS1)表达,四甲基偶氮唑蓝(MTT)法和集落形成实验检测细胞增殖能力,流式细胞术检测细胞凋亡和周期分布,蛋白质印记检测Ki67、半胱氨酸蛋白酶3(Pro-caspase3)和裂解的Caspase3(Cleaved-caspase3)表达。将HIF1A-AS1过表达质粒转染SW480细胞,采用上述方法检测过表达HIF1A-AS1对罗仙子提取物处理的SW480细胞增殖、凋亡和周期分布的影响。结果:低、高剂量的罗仙子提取物能降低SW480细胞中HIF1A-AS1表达水平,降低细胞存活率、克隆形成数和S期细胞比例,升高G_(0)~G_(1)期细胞比例和细胞凋亡率,下调Ki67和Pro-caspase3蛋白表达,上调Cleaved-caspase3蛋白表达(P<0.05)。过表达HIF1A-AS1能够逆转罗仙子提取物对SW480细胞增殖、凋亡、周期分布以及Ki67、Pro-caspase3和Cleaved-caspase3蛋白表达的影响,恢复SW480细胞的增殖能力和凋亡水平(P<0.05)。结论:一定浓度的罗仙子提取物可抑制结直肠癌细胞增殖,诱导G_(0)~G_(1)期阻滞和凋亡,具有防治结肠癌的应用前景,其机制可能与下调HIF1A-AS1表达有关。Objective:To study the effect of housefly maggots extracts on proliferation,apoptosis and cycle distribution of colorectal cancer cells and its possible mechanism.Methods:The colon cancer cells(SW480)were treated with medium,low and high doses of housefly maggots extracts for 24 h,real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of long non-coding RNA(lncRNA)hypoxia inducer 1 antisense RNA1(HIF1A-AS1),methyl thiazolyl tetrazolium(MTT)method and colony formation experiment were used to detect the cell proliferation ability,flow cytometry was used to detect apoptosis and cycle distribution,Western blot was used to detect the expression of Ki67,cysteine??proteinase 3(Pro-caspase3)and Cleaved-caspase3.The HIF1A-AS1 overexpression plasmid was transfected into SW480 cells,and the effects of HIF1A-AS1 overexpression on the proliferation,apoptosis and cycle distribution of SW480 cells treated with housefly maggots extracts were detected by the above method.Results:Low and high dose of housefly maggots extracts decreased the expression of HIF1A-AS1,Ki67 and Pro-caspase3 of SW480 cells,decreased cell survival rate,the number of clone formation and the S phase proportion,as well as increased G0-G1 phase proportion,apoptosis rate and Cleared-caspase3 protein expression level(P<0.05).Overexpression of HIF1A-AS1 can reverse the effects of housefly maggots extracts on cell proliferation,apoptosis,cycle distribution and expression of Ki67,Pro-caspase3 and Cleared-caspase3 proteins,and restore the proliferation and apoptosis levels of SW480 cells(P<0.05).Conclusions:A certain concentration of housefly maggots extracts can inhibit the proliferation of colorectal cancer cells,induce G0-G1 phase arrest and apoptosis,and has the application prospect in prevention of colon cancer.The mechanism may be related to the down-regulation of HIF1A-AS1 expression.

关 键 词：罗仙子提取物 结直肠癌 HIF1A-AS1 增殖 凋亡 周期分布 

分 类 号：R735.3[医药卫生—肿瘤]