丹参酮ⅡA联合CASC2对甲状腺癌细胞增殖、凋亡、迁移、侵袭的影响固有淋巴细胞在脂质代谢中的研究进展

Effects of TanshinoneⅡA Combined with CASC2 on Proliferation,Apoptosis,Migration and Invasion of Thyroid Cancer Cells

为了研究丹参酮ⅡA联合长链非编码RNA(lnc RNA)癌易感性候选基因2(CASC2)对甲状腺癌细胞增殖、凋亡、迁移、侵袭的影响,该研究采用实时荧光定量PCR(q RT-PCR)检测CASC2在甲状腺癌组织中的表达。将甲状腺癌SW579细胞分为pc DNA3.1组(转染pc DNA3.1质粒),pc DNA3.1-CASC2组(转染pc DNA3.1-CASC2质粒),con组(用与丹参酮ⅡA等量的二甲基亚砜处理),药物-1、2、3、4组(分别用1、2、4、8μg/m L丹参酮ⅡA处理),药物-4+pc DNA3.1组(转染pc DNA3.1质粒且用8μg/m L丹参酮ⅡA处理),药物-4+pc DNA3.1-CASC2组(转染pc DNA3.1-CASC2质粒且用8μg/m L丹参酮ⅡA处理)。分别用细胞计数试剂盒(CCK-8)和平板克隆检测细胞存活与克隆形成;流式细胞术检测细胞凋亡;Transwell检测细胞迁移、侵袭;蛋白质印迹法(Western blot)检测蛋白P21、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、E-钙黏蛋白(E-cadherin)和基质金属蛋白酶-2(MMP-2)的表达。结果显示,与癌旁组织相比,甲状腺癌组织中的CASC2表达量显著降低(P<0.05)。过表达CASC2明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数和MMP-2蛋白表达量,显著提高细胞凋亡率、P21、Caspase-3、E-cadherin蛋白表达量(P<0.05)。丹参酮ⅡA明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白水平,显著提高细胞凋亡率、P21、Caspase-3、E-cadherin蛋白表达水平,且均呈浓度依赖性(P<0.05)。丹参酮ⅡA联合CASC2明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白表达量,显著提高细胞凋亡率、P21、Caspase-3和E-cadherin蛋白水平(P<0.05)。因此,丹参酮ⅡA联合CASC2可以抑制甲状腺癌细胞的增殖、迁移和侵袭,以及诱导细胞凋亡。

To study the effect of tanshinone IIA combined with lnc RNA(long non-coding RNA)CASC2(cancer susceptibility candidate 2)on the proliferation,apoptosis,migration and invasion of thyroid cancer cells,this study detected the expression of CASC2 in thyroid cancer tissues by q RT-PCR(real-time fluorescent quantitative PCR).Thyroid cancer SW579 cells were divided into pc DNA3.1 group(transfected with pc DNA3.1 plasmid),pc DNA3.1-CASC2 group(transfected with pc DNA3.1-CASC2 plasmid),con group(treated with dimethyl sulfoxide in the same amount as tanshinone IIA),medicine-1,2,3,4 groups(separately treated with 1,2,4,8μg/m L tanshinone IIA),medicine-4+pc DNA3.1 group(transfected with pc DNA3.1 plasmid and treated with 8μg/m L tanshinone IIA),medicine-4+pc DNA3.1-CASC2 group(transfected with pc DNA3.1-CASC2 plasmid and treated with 8μg/m L tanshinone IIA).CCK-8(cell counting kit-8)and plate clones separately were used to detect cell survival and colony formation.Cell apoptosis was examined by flow cytometry.Transwell was applied to determine cell migration and invasion,and protein P21,Caspase-3(Cysteinyl aspartate specific proteinase 3),E-cadherin and MMP-2(matrix metalloproteinase-2)expression were analyzed by Western blot.The results showed that,compared with adjacent tissues,the expression of CASC2 in thyroid cancer tissues was significantly reduced(P<0.05).Overexpression of CASC2 obviously decreased the survival rate,colony formation,the number of migrating cells,the number of invading cells,and MMP-2 protein expression in SW579 cells,while markedly increased the apoptosis rate,as well as P21,Caspase-3,and Ecadherin protein expression(P<0.05).Tanshinone IIA dramatically reduced the survival rate,the number of colony formation,the number of migrating cells,the number of invading cells,and the level of MMP-2 protein in SW579 cells,while distinctly improved the rate of apoptosis,and the expression levels of P21,Caspase-3,and E-cadherin proteins,which were all concentration-dependent(P<0.05).Tanshinone IIA combined with CASC2 remarkably reduced the survival rate,colony formation,the number of migrating cells,the number of invading cells,and MMP-2 protein expression in SW579 cells,while greatly enhanced the apoptosis rate,as well as P21,Caspase-3,and E-cadherin protein levels(P<0.05).So it can be concluded that,tanshinone IIA combined with CASC2 can inhibit the proliferation,migration and invasion of thyroid cancer cells,and induce cell apoptosis.