使君子醇提物下调TPT1-AS1抑制肝癌细胞增殖、侵袭迁移

Inhibition of the Proliferation,Invasion and Migration of Liver Cancer Cells by Down-regulating TPT1-AS1 of Alcohol Extract of Quisqualis Indica L.

目的:考察使君子醇提物是否通过下调长链非编码RNA(lnc RNA)TPT1-AS1来抑制肝癌细胞的增殖、侵袭和迁移。方法:采用噻唑蓝(MTT)法检测使君子醇提物(10,50,100μg·ml-1)作用于肝癌HCC9204细胞的存活率,Transwell评估HCC9204细胞侵袭迁移,实时荧光定量PCR测定HCC9204细胞中TPT1-AS1表达,免疫印迹实验(Western blot)分析细胞核相关抗原Ki-67(Ki-67)、神经型钙黏蛋白(N-cadherin)、上皮型钙黏蛋白(E-cadherin)蛋白的表达。在HCC9204细胞中转染TPT1-AS1小干扰RNA si-TPT1-AS1,或转染TPT1-AS1过表达质粒pc DNA-TPT1-AS1并使用100μg·ml-1使君子醇提物处理,评价其对细胞的增殖、侵袭和迁移的影响。结果:50和100μg·ml-1使君子醇提物显著减少HCC9204细胞的存活率、迁移细胞数、侵袭细胞数、TPT1-AS1表达水平、Ki67、N-cadherin的蛋白表达水平,显著增加E-cadherin的蛋白表达水平(P<0.05)。敲减TPT1-AS1显著降低HCC9204细胞的存活率、迁移细胞数、侵袭细胞数、Ki67、N-cadherin的蛋白表达水平,显著提高Ecadherin的蛋白表达水平(P<0.05)。TPT1-AS1过表达逆转了使君子醇提物抑制HCC9204细胞存活、侵袭、迁移、Ki67、N-cadherin蛋白表达的作用,并逆转了其促进HCC9204细胞E-cadherin蛋白表达的作用。结论:使君子醇提物通过下调TPT1-AS1的表达,抑制肝癌细胞的增殖、侵袭和迁移能力。

Objective:To investigate whether Quisqualis indica L.extract can inhibit the proliferation,invasion and migration of liver cancer cells by down-regulating long-chain non-coding RNA(lnc RNA)TPT1-AS1.Methods:The thiazole blue(MTT)method was used to detect the survival rate of hepatocellular carcinoma HCC9204 cells treated with Quisqualis indica L.extract(10,50,100μg·ml-1).Transwell was applied to evaluate the invasion and migration of HCC9204 cells.Real-time fluorescent quantitative PCR was used to determine the expression of TPT1-AS1 in HCC9204 cells.Western blot was used to analyze nuclear-associated antigen Ki-67(Ki-67),neuronal cadherin(N-cadherin)and epithelial cadherin(E-cadherin)protein expressions.TPT1-AS1 small interfering RNA si-TPT1-AS1 or TPT1-AS1 overexpression plasmid pc DNA-TPT1-AS1 was transfected into HCC9204 cells,followed by the treatment with 100μg·ml-1 Quisqualis indica L.extract to evaluate the proliferation,invasion and migration of cells.Results:Quisqualis indica L.extract at 50 and 100μg·ml-1 significantly decreased the survival rate,migration number,invasion cell number,TPT1-AS1 expression level,Ki67 and N-cadherin protein expression levels of HCC9204 cells,and greatly increased the protein expression level of E-cadherin(P<0.05).Knocking down TPT1-AS1 dramatically reduced the survival rate of HCC9204 cells,the number of migrating cells,the number of invasive cells,the protein expression levels of Ki67 and N-cadherin,while remarkably increased the protein expression level of E-cadherin(P<0.05).Overexpression of TPT1-AS1 reversed the above effects of Quisqualis indica L.extract in HCC9204 cells.Conclusion:Quisqualis indica L.extract can inhibit the proliferation,invasion and migration of liver cancer cells by down-regulating the expression of TPT1-AS1.