南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (3): 447-452.doi: 10.12122/j.issn.1673-4254.2021.03.19

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SPOP上调c-Jun蛋白表达并促进肾癌细胞的增殖和侵袭

吴林慧，余 珂，崔 悦，朱雪莲，杨 正，马 佳   

  • 出版日期:2021-03-20 发布日期:2021-04-06

Speckle-type POZ protein up-regulates c-Jun protein expression and promotes proliferation and invasion of renal carcinoma cells

  • Online:2021-03-20 Published:2021-04-06

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摘要： 目的 探讨SPOP对肾癌细胞增殖、凋亡、迁移和侵袭等的影响及潜在的作用机制。方法 体外培养肾癌细胞株786-O、 A704、Caki-2，对照组（EV）和过表达组（SPOP）质粒采用脂质体法转染至上述肾癌细胞株；分别利用克隆形成实验及MTT法检测SPOP基因对肾癌细胞株增殖的影响；采用TUNEL法检测SPOP基因对肾癌细胞株凋亡的影响；利用创伤愈合实验和Transwell实验检测SPOP基因对肾癌细胞株迁移和侵袭能力的作用；运用Real-time PCR和Western blotting技术检测转染后肾癌细胞株SPOP和c-Jun的 mRNA水平以及相关蛋白的表达情况。结果 在786-O、A704、Caki-2细胞上调 SPOP的表达能促进肾癌细胞的增殖、迁移和侵袭（P<0.05）；与对照组相比，SPOP组的肾癌细胞株786-O、Caki-2的凋亡率较对照组相对减少（P< 0.05）；Real-time PCR结果显示，c-Jun mRNA的表达水平未发生改变，但Western blotting结果显示，SPOP组肾癌细胞株786-O、Caki-2的SPOP蛋白表达明显高于对照组（P<0.05），c-Jun蛋白的表达也明显高于对照组（P<0.05）。结论 SPOP能促进肾癌细胞的增殖、迁移和侵袭能力，抑制肾癌细胞的凋亡，潜在的机制可能为促进c-Jun的表达。

关键词: SPOP；c-Jun；肾癌；增殖；迁移；侵袭；凋亡

Abstract: Objective To investigate the effect of speckle-type POZ protein (SPOP) on proliferation, apoptosis, migration and invasion of renal cell carcinoma (RCC) and explore the potential mechanisms. Methods Renal carcinoma cell lines (786-O, A704, and Caki-2) cultured in vitro were transfected with a SPOP-overexpressing plasmid, and the changes in proliferation of the cells were detected using colony formation and MTT assay; TUNEL assay was used to assess apoptosis of the cells. The changes in migration and invasion abilities of the cells were examined using wound healing assay and Transwell assay. The mRNA and protein levels of SPOP and c-Jun in the transfected cells were measured using real-time PCR and Western blotting. Results SPOP over-expression obviously promoted the proliferation, migration and invasion of 786-O, A704 and Caki-2 cells (P<0.05). Compared with the control cells, 786-o and Caki-2 cells over-expressing SPOP exhibited significantly lowered apoptosis rates (P<0.05). The results of real-time PCR demonstrated that the transfected cells did not show obvious changes in the mRNA level of c-Jun, but the protein expressions of SPOP and c-jun increased significantly as shown by Western blotting (P<0.05). Conclusion SPOP can promote proliferation, migration, and invasion and suppress apoptosis of renal carcinoma cells possibly by promoting the expression of c-Jun.

Key words: renal carcinoma; speckle-type POZ protein; c-Jun; proliferation; migration; invasion; apoptosis