南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (4): 549-554.doi: 10.12122/j.issn.1673-4254.2021.04.10

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siRNA 干扰 CTHRC1 可体外抑制甲状腺乳头状癌 TPC-1细胞的增殖并诱导凋亡

唐振宁，丁小云，秦少杰，张朝林   

  • 出版日期:2021-04-20 发布日期:2021-04-30

Effects of RNA interference of CTHRC1 on proliferation and apoptosis of thyroid papillary cancer TCP-1 cells in vitro

  • Online:2021-04-20 Published:2021-04-30

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摘要： 目的 探讨胶原三股螺旋重复蛋白1（CTHRC1）甲状腺乳头状癌TPC-1细胞增殖、凋亡的影响。方法 成功筛选干扰甲状腺乳头状癌TPC-1细胞CTHRC1表达的siRNA，以脂质体转染法将siRNA转染甲状腺乳头状癌TPC-1细胞，采用real-time PCR和Western blot检测转染后TCP-1细胞中CTHRC1mRNA和蛋白表达的变化。实验分组：将筛选的干扰序列转染TPC-1细胞作为干扰组（si-CTHRC1组），将随机阴性对照序列转染的TPC-1细胞作为阴性对照组（si-NC组），以不做任何处理TPC-1细胞为空白组（WT组）。采用CCK-8实验检测细胞增殖；流式细胞术AV/PI双染观察各组TCP-1细胞凋亡的变化；采用Western blot法检测各组 TPC-1 细胞中含剪切型半胱氨酸的天冬氨酸蛋白水解酶-3（c-Caspase-3）蛋白、剪切型多聚 ADP 核糖聚合酶 1 （c-PARP1）蛋白表达水平以及磷酸化细胞外信号调节激酶 1/2(ERK1/2)表达水平。结果 将筛选出明显抑制 TPC-1 细胞CTHCR1表达的siRNA转染细胞后，TPC-1细胞中CTHRC1 的mRNA和蛋白表达水平降低（P<0.05）。与WT组和si-NC组相比，si-CTHRC1组的细胞活降低（P<0.05）；与对照组相比，si-CTHRC1组的TPC-1细胞细胞凋亡率升高，Western blot实验发现：si-CTHRC1组的c-caspase-3、c-PARP1蛋白表达水平均升高（P<0.05）；si-CTHRC1组ERK1/2蛋白的磷酸化水平增加。结论 干扰CTHRC1能够抑制TPC-1细胞的增殖并诱导凋亡，此过程可能通过激活ERK1/2信号通路介导。

关键词: 甲状腺癌；CTHRC1；增殖；凋亡；siRNA干扰

Abstract: Objective To explore the role of CTHRC1 in regulating the proliferation and apoptosis of papillary thyroid cancer cells. Methods Papillary thyroid cancer TPC-1 cells were transfected with a small interfering RNA (siRNA) targeting CTHRC1, with the cells transfected with a scrambled sequence as the negative control. The changes in cell proliferation and apoptosis were assessed using cell counting kit-8 (CCK-8) and flow cytometry with AV/PI double staining, respectively. The expression of c-caspase-3, c-PARP1 and phosphorylation of ERK1/2 in the cells were examined with Western blotting. Results Transfection with the siRNA sequence significantly decreased the mRNA and protein levels of CTHRC1 in TCP-1 cells (P<0.05). Compared with blank and negative control cells, TCP-1 cells with RNA interference of CTHRC1 showed significantly lowered proliferative activity and enhanced cell apoptosis (P<0.05) with significantly increased expressions of c-caspase-3 and c-PARP1 and phosphorylation of ERK1/2 (P<0.05). Conclusion RNA interference of CTHRC1 promotes the proliferation and inhibits apoptosis of papillary thyroid cancer cells possibly by activating the ERK1/2 pathway

Key words: thyroid carcinoma; CTHRC1; proliferation; apoptosis; siRNA interference