CDC25A与IL-6在肝癌HepG2细胞中的表达及相互作用研究
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摘要
目的:本研究旨在进一步研究CDC25A基因对肝癌HepG2细胞增殖、侵袭和转移的影响,探讨CDC25A与IL-6在肝癌中是否存在相互作用。方法:用慢病毒CDC25A-shRNA转染HepG2细胞以特异性阻断CDC25A的表达为实验组(KD组),用慢病毒阴性shRNA转染作为阴性对照组(NC组),用常规培养的HepG2细胞为空白对照组(Control组)。采用实时荧光定量PCR、Western blot检测CDC25A、IL-6的mRNA及蛋白的表达水平。结果:实验组IL-6 mRNA的表达水平明显低于阴性对照组和空白对照组,差异具有统计学意义(P<0.05)。IL-6的蛋白表达水平明显低于阴性对照组和空白对照组,差异具有统计学意义(P<0.05)。结论:沉默肝癌HepG2细胞中的CDC25A基因,IL-6的表达下调。CDC25A可能通过调节IL-6的表达,在肝癌的发生发展及侵袭过程中发挥作用。
Objective: To further investigate the effect of CDC25 A gene on proliferation, invasion and metastasis of hepatocellular carcinoma HepG2 cells, and to explore whether CDC25 A and IL-6 interact in liver cancer. Methods: HepG2 cells were transfected with lentiviral CDC25 A-shRNA to specifically block the expression of CDC25 A in the experimental group(KD group), transfected with lentiviral-negative shRNA as a negative control group(NC group), and the conventionally cultured HepG2 cells were blank control group(Control group). Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expression levels of CDC25 A and IL-6. Results: The expression level of IL-6 mRNA in the experimental group was significantly(P<0.05) lower than that of the negative control group and the blank control group. The protein expression level of IL-6 in the experimental group was significantly lower than that of the negative control group and the blank control group. The difference was statistically significant(P<0.05). Conclusions: The CDC25 A gene in hepatoma HepG2 cells was silenced and the expression of IL-6 was down-regulated. CDC25 A may play a role in the development and invasion of liver cancer by regulating the expression of IL-6.
Objective: To further investigate the effect of CDC25 A gene on proliferation, invasion and metastasis of hepatocellular carcinoma HepG2 cells, and to explore whether CDC25 A and IL-6 interact in liver cancer. Methods: HepG2 cells were transfected with lentiviral CDC25 A-shRNA to specifically block the expression of CDC25 A in the experimental group(KD group), transfected with lentiviral-negative shRNA as a negative control group(NC group), and the conventionally cultured HepG2 cells were blank control group(Control group). Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expression levels of CDC25 A and IL-6. Results: The expression level of IL-6 mRNA in the experimental group was significantly(P<0.05) lower than that of the negative control group and the blank control group. The protein expression level of IL-6 in the experimental group was significantly lower than that of the negative control group and the blank control group. The difference was statistically significant(P<0.05). Conclusions: The CDC25 A gene in hepatoma HepG2 cells was silenced and the expression of IL-6 was down-regulated. CDC25 A may play a role in the development and invasion of liver cancer by regulating the expression of IL-6.
引文
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2 Bray F,Ferlay J,Soerjomataram I,et al.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J].CA Cancer J Clin,2018,68(6):394-424.
3 Dozier C,Mazzolini L,Cénac C,et al.CyclinD-CDK4/6 complexes phosphorylate CDC25A and regulate its stability[J].Oncogene,2017,36(26):3781-3788.
4 Sadeghi H,Golalipour M,Yamchi A,et al.CDC25A pathway toward tumorigenesis:Molecular targets of CDC25A in cell-cycle regulation[J].J Cell Biochem,2019,120(3):2919-2928.
5 王红雷.CDC25A在肝细胞癌中的表达及意义[J].山东医药,2014,54(25):45-46.
6 李薇.RNA干扰技术沉默CDC25a基因对人肝癌HepG2细胞增殖和裸鼠移植瘤的影响[D].南宁:广西医科大学,2015.
7 Sarkis M,Miteva MA,Dasso Lang MC,et al.Insights into the interaction of high potency inhibitor IRC-083864 with phosphatase CDC25[J].Proteins,2017,85(4):593-601.
8 Newell P,Villanueva A,Llovet JM.Molecular targeted therapies in hepatocellular carcinoma:from pre-clinical models to clinical trials[J].J Hepatol,2008,49(1):1-5.
9 Jung IH,Choi JHK,Chung YY,et al.Predominant activation of JAK/STAT3 pathway by interleukin-6 is implicated in Hepatocarcinogenesis12[J].Neoplasia,2015,17(7):586-597.
10 He GB,Dhar D,Nakagawa H,et al.Identification of liver cancer progenitors whose malignant progression depends on autocrine IL-6 signaling[J].Cell,2013,155(2):384-396.
11 Sur S,Agrawal DK.Phosphatases and kinases regulating CDC25 activity in the cell cycle:clinical implications of CDC25 overexpression and potential treatment strategies[J].Mol Cell Biochem,2016,416(1/2):33-46.
12 de Gooijer MC,van den Top A,Bockaj I,et al.The G2 checkpoint-a node-based molecular switch[J].FEBS Open Bio,2017,7(4):439-455.
13 Tsuchiya Y,Murai S,Yamashita S.Dual inhibition of Cdc2 protein kinase activation during apoptosis in Xenopus egg extracts[J].FEBS J,2015,282(7):1256-1270.
14 Qin HZ,Liu WF.MicroRNA-99a-5p suppresses breast cancer progression and cell-cycle pathway through downregulating CDC25A[J].J Cell Physiol,2019,234(4):3526-3537.
15 Wang XQ,Zhu YQ,Lui KS,et al.Aberrant polo-like kinase 1-Cdc25A pathway in metastatic hepatocellular carcinoma[J].Clin Cancer Res,2008,14(21):6813-6820.
16 Xu XD,Yamamoto H,Liu GX,et al.CDC25A inhibition suppresses the growth and invasion of human hepatocellular carcinoma cells[J].Int J Mol Med,2008,21(2):145-152.
17 Lu XX,Sun W,Tang YP,et al.Identification of key genes in hepatocellular carcinoma and validation of the candidate gene,cdc25a,using gene set enrichment analysis,meta-analysis and cross-species comparison[J].Mol Med Rep,2016,13(2):1172-1178.
18 王华敏.miR-197与IL-6/STAT3信号通路相互调控在肝癌生长的作用及机制研究[D].杭州:浙江大学,2015.
19 Ma HY,Yan D,Wang YN,et al.Bazedoxifene exhibits growth suppressive activity by targeting interleukin-6/glycoprotein 130/signal transducer and activator of transcription 3 signaling in hepatocellular carcinoma[J].Cancer Sci,2019,110(3):950-961.
20 Liu XQ,Zong W,Li TL,et al.The E3 ubiquitin ligase APC/CCdh1 degrades MCPH1 after MCPH1-βTrCP2-Cdc25A-mediated mitotic entry to ensure neurogenesis[J].EMBO J,2017,36(24):3666-3681.
21 Liu XM,Zhang AP,Xiang JX,et al.MiR-451 Acts as a suppressor of angiogenesis in hepatocellular carcinoma by targeting the IL-6R-STAT3 pathway[J].Oncol Rep,2016,36(3):1385-1392.