摘要
目的探究过表达低氧诱导因子-1(HIF-1α)对三阴性乳腺癌MDA-MB-231细胞化疗耐药及其对体内外增殖机制的影响。方法2020年1—12月于海南省肿瘤医院进行实验。(1)MDA-MB-231细胞培养后随机分为3组,空白对照组不进行转染,pEGFP-HIF-1α组、pEGFP-C1组,分别将pEGFP-HIF-1α质粒及pEGFP-C1空载体质粒转染入细胞中。采用荧光定量PCR与Western-blot检测转染后HIF-1α、VEGF的mRNA和蛋白表达,CCK-8法检测细胞药物敏感性与细胞生长抑制率,Annexin V-FITC/PI双染法检测细胞凋亡。(2)将20只雌性BALB/c裸鼠随机数字表法分为C1组和HIF-1α组,每组10只,HIF-1α组裸鼠左侧乳垫处注射转染pEGFP-HIF-1α的MDA-MB-231细胞,C1组裸鼠左侧乳垫处注射转染pEGFP-C1的MDA-MB-231细胞,1周后观察裸鼠成瘤状况,于4周后处死裸鼠,取肿瘤组织HE染色检测组织病理学变化,免疫组织化学染色检测Ki-67与VEGF的表达。结果与空白对照组比较,pEGFP-HIF-1α组细胞中HIF-1αmRNA和蛋白相对表达量均显著升高(t/P=36.027/0.000、25.432/0.000),VEGF mRNA和蛋白相对表达量均显著升高(t/P=24.551/0.000、19.876/0.000);顺铂IC50值显著增加(P<0.05),顺铂处理培养72、96 h后细胞生长抑制率显著下降(P<0.05),细胞凋亡率显著降低(t/P=56.437/0.000);与C1组比较,HIF-1α组裸鼠在1、2、3、4周时肿瘤体积显著增大(t/P=79.987/0.000、120.547/0.000、89.152/0.000、80.984/0.000),肿瘤组织中肿瘤细胞排列较为密集,Ki-67、VEGF的阳性表达率显著增加(t/P=83.436/0.000、67.817/0.000)。结论过表达HIF-1α会降低MDA-MB-231细胞对顺铂的敏感性,抑制细胞凋亡,促进裸鼠三阴性乳腺癌肿瘤组织的生长,其机制可能与促进VEGF表达有关。
Objective To explore the effect of overexpression of hypoxia-inducible factor-1(HIF-1α)on chemotherapy resistance of triple-negative breast cancer MDA-MB-231 cells and its proliferation mechanisms in vivo and in vitro.Method The experiment will be conducted in Hainan Cancer Hospital from January to December 2020.(1)MDA-MB-231 cells were randomly divided into 3 groups after culture.The pEGFP HIF-1αplasmid(pEGFP HIF-1αgroup)and pEGFP-C1 empty vector plasmid(pEGFP-C1 group)were transfected into the cells.The blank control group cells were not transfected.Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of HIF-1αand VEGF after transfection,CCK-8 method was used to detect cell drug sensitivity and cell growth inhibition rate,and Annexin V FITC/PI double staining method was used to detect cell apoptosis.(2)Divide 20 female BALB/c nude mice into C1 group and HIF-1αgroup by random number table,with 10 mice in each group.HIF-1αgroup nude mice were injected with pEGFP HIF-1α-transfected MDA-MB-231 cells on the left mammary pad,and C1 group nude mice were injected with pEGFP-C1 transfected MDA-MB-231 cells on the left mammary pad.One week later,the tumorigenesis status of nude mice was observed,and the nude mice were sacrificed 4 weeks later.HE staining was used to detect tumor histopathological tissue changes,and immunohistochemical staining was used to detect the expression of Ki-67 and VEGF.Results Compared with the blank control group,the relative expression of HIF-1αmRNA and protein in the cells of the pEGFP-HIF-1αgroup was significantly increased(t/P=36.027/0.000,25.432/0.000),and the relative expression of VEGF mRNA and protein in the cells were significantly increased(t/P=24.551/0.000,19.876/0.000).and the IC50 value of cisplatin was significantly increased(P<0.05).After treatment and culture for 72 and 96 h,the cell growth inhibition rate was significantly decreased(P<0.05),the cell apoptosis rate was significantly reduced(t/P=56.437/0.000),Compared with th
作者
王亮
黄琰菁
娄娜娜
吴焕良
Wang Liang;Huang Yanjing;Lou Nana;Wu Huanliang(Department of Radiotherapy, Hainan Cancer Hospital, Hainan Province, Haikou 570311,China)
出处
《疑难病杂志》
CAS
2021年第10期997-1002,共6页
Chinese Journal of Difficult and Complicated Cases
基金
海南省自然科学基金面上项目(819MS117)。
关键词
三阴性乳腺癌
低氧诱导因子-1Α
化疗敏感性
增殖
裸鼠
Triple negative breast cancer
Hypoxia inducible factor 1α
Chemotherapy sensitivity
Proliferation
Nude mice
