CUL4A 通过调控 Hippo 通路促进结肠癌增殖并抑制凋亡。
CUL4A promotes proliferation and inhibits apoptosis of colon cancer cells via regulating Hippo pathway.
机构信息
Department of Gastroenterology, Yantaishan Hospital, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Oct;24(20):10518-10525. doi: 10.26355/eurrev_202010_23404.
OBJECTIVE
The aim of this study was to investigate the effect of cullin 4A (CUL4A) on the proliferation and apoptosis of colon cancer (CC) cells, and to elucidate its regulatory relationship with the Hippo pathway.
PATIENTS AND METHODS
Paired CC tissues and adjacent normal tissues were obtained from patients. CC cells were isolated and cultured in vitro. CUL4A was interfered by small interfering ribonucleic acid (siRNA) (siR-CUL4A group) or overexpressed by overexpression vector (CUL4A-Vector group), with negative control (NC)-CUL4A or CUL4A -NC as the control group. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of CUL4A in CC tissues and cells. The proliferative ability of cells was detected by cell counting kit-8 (CCK-8) assay. Flow cytometry was applied to measure the apoptosis of cells in each group. Western blotting (WB) was conducted to determine the protein expression of CUL4A. In addition, the proliferative ability was examined in vivo through subcutaneous injection of cells into nude mice.
RESULTS
QRT-PCR showed that CUL4A was highly expressed in 66.67% of CC samples (p<0.01). In vivo and in vitro proliferative ability was significantly reduced in siR-CUL4A group (p<0.01), whereas the apoptosis rate was promoted (p<0.01). However, in vivo and in vitro proliferative ability increased significantly in CUL4A-Vector group (p<0.01), while the apoptosis rate was reduced (p<0.01). The protein expressions of MST1, LATS1 and p-YAP were significantly up-regulated in siR-CUL4A group (p<0.01), while they were remarkably down-regulated in CUL4A-Vector group (p<0.05, p<0.01).
CONCLUSIONS
CUL4A is highly expressed in CC and promotes the proliferation and inhibits the apoptosis of CC cells by regulating the Hippo pathway.
目的
本研究旨在探讨 CUL4A（细胞周期蛋白 4A）对结肠癌（CC）细胞增殖和凋亡的影响，并阐明其与 Hippo 通路的调控关系。
方法
收集患者配对的 CC 组织和相邻正常组织，体外分离培养 CC 细胞。用小干扰 RNA（siRNA）干扰 CUL4A（siR-CUL4A 组）或过表达载体过表达 CUL4A（CUL4A-Vector 组），以阴性对照（NC）-CUL4A 或 CUL4A-NC 作为对照组。采用实时定量聚合酶链反应（qRT-PCR）检测 CC 组织和细胞中 CUL4A 的表达水平。用细胞计数试剂盒-8（CCK-8）法检测细胞的增殖能力。流式细胞术检测各组细胞的凋亡情况。Western blot（WB）法检测 CUL4A 的蛋白表达。此外，通过将细胞皮下注射到裸鼠体内来检测体内的增殖能力。
结果
qRT-PCR 显示，CUL4A 在 66.67%的 CC 样本中高表达（p<0.01）。siR-CUL4A 组体内和体外的增殖能力明显降低（p<0.01），而凋亡率升高（p<0.01）。然而，CUL4A-Vector 组体内和体外的增殖能力显著增加（p<0.01），而凋亡率降低（p<0.01）。siR-CUL4A 组 MST1、LATS1 和 p-YAP 的蛋白表达明显上调（p<0.01），而 CUL4A-Vector 组则明显下调（p<0.05，p<0.01）。
结论
CUL4A 在 CC 中高表达，通过调节 Hippo 通路促进 CC 细胞的增殖并抑制其凋亡。
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