Tet methylcytosine dioxygenase 2(TET2)acts as an antioncogene that is investigated in different cancers.But the effects of TET2 in renal cell cancer(RCC)is still known little.Here,quantitative real-time PCR(qRT-PCR),W...Tet methylcytosine dioxygenase 2(TET2)acts as an antioncogene that is investigated in different cancers.But the effects of TET2 in renal cell cancer(RCC)is still known little.Here,quantitative real-time PCR(qRT-PCR),Western blot,and immunofluorescence were performed to exam gene and protein expression.Cell proliferation was measured using Cell Counting Kit-8(CCK-8).Transwell assay was performed to detect cell metastasis viability.Flow cytometry was performed to analyze the cell cycle and cell apoptosis.The effects of TET2 on RCC growth in vivo was analyzed using a mouse xenograft model.We found that TET2 and miR-200c were decreased in RCC tissues,and hypermethylation of miR-200c promoter was found.Overexpression of TET2 promoted miR-200c expression by reducing miR-200c promoter methylation.Additionally,overexpression of TET2 or miR-200c suppressed cell growth and metastasis.Also,knockdown of miR-200c could moderate TET2 mediated cell growth inhibition.Furthermore,we found miR-200c directly regulates Stearoyl-CoA desaturase(SCD)gene expression.Moreover,in vivo experiment results confirmed that TET2 inhibited tumor growth.In conclusion,TET2 acts as an antioncogene in RCC by regulating the miR-200c-SCD axis and providing a potential target for RCC diagnosis and treatment.展开更多
目的应用hIAP-2特异性siRNA干扰肾癌GRC-1细胞hIAP-2基因的表达,探讨siRNA-hIAP-2联合姜黄素对肾癌GRC-1细胞增殖和凋亡的影响。方法体外化学合成针对hIAP-2的siRNA,通过阳离子脂质体将siRNA-hIAP-2转染肾癌GRC-1细胞,实时荧光定量PCR检...目的应用hIAP-2特异性siRNA干扰肾癌GRC-1细胞hIAP-2基因的表达,探讨siRNA-hIAP-2联合姜黄素对肾癌GRC-1细胞增殖和凋亡的影响。方法体外化学合成针对hIAP-2的siRNA,通过阳离子脂质体将siRNA-hIAP-2转染肾癌GRC-1细胞,实时荧光定量PCR检测hIAP-2 m RNA表达水平,免疫印迹检测其蛋白表达水平,MTT法及流式细胞术分别测定siRNA-hIAP-2联合姜黄素处理后对肾癌GRC-1细胞增殖及凋亡的影响。结果靶向hIAP-2的siRNA可以有效抑制肾癌GRC-1细胞的hIAP-2的表达,siRNA-hIAP-2和姜黄素均可抑制GRC-1细胞增殖并诱导其一定程度的凋亡,当联合应用靶向hIAP-2的siRNA和姜黄素时,上述作用显著增强(P<0.05)。结论体外化学合成的siRNA-hIAP-2可以有效抑制肾癌GRC-1细胞的hIAP-2的表达,并能显著增强姜黄素诱导肾癌细胞GRC-1凋亡的敏感度,提示hIAP-2可作为姜黄素治疗肾癌的有效增敏靶点。展开更多
The tumor suppressor p53 transactivates the expression of multiple genes to exert its multifaceted functions and ultimately maintains genome stability.Thus,cancer cells develop various mechanisms to diminish p53 expre...The tumor suppressor p53 transactivates the expression of multiple genes to exert its multifaceted functions and ultimately maintains genome stability.Thus,cancer cells develop various mechanisms to diminish p53 expression and bypass the cell cycle checkpoint.In this study,we identified the gene encoding RNAbinding protein cytoplasmic polyadenylation element-binding protein 2(CPEB2)as a p53 target.In turn,CPEB2 decreases p53 messenger RNA stability and translation to fine-tune p53 level.Specifically,we showed that CPEB2 binds the cytoplasmic polyadenylation elements in the p5330-untranslated region,and the RNA recognition motif and zinc finger(ZF)domains of CPEB2 are required for this binding.Furthermore,we found that CPEB2 was upregulated in renal cancer tissues and promotes the renal cancer cell proliferation and migration.The oncogenic effect of CPEB2 is partially dependent on negative feedback regulation of p53.Overall,we identify a novel regulatory feedback loop between p53 and CPEB2 and demonstrate that CPEB2 promotes tumor progression by inactivating p53,suggesting that CPEB2 is a potential therapeutic target in human renal cancer.展开更多
基金supported by Grants from the Nature Science Foundation of Fujian,China(Nos.2010J01372,2015J01571).
文摘Tet methylcytosine dioxygenase 2(TET2)acts as an antioncogene that is investigated in different cancers.But the effects of TET2 in renal cell cancer(RCC)is still known little.Here,quantitative real-time PCR(qRT-PCR),Western blot,and immunofluorescence were performed to exam gene and protein expression.Cell proliferation was measured using Cell Counting Kit-8(CCK-8).Transwell assay was performed to detect cell metastasis viability.Flow cytometry was performed to analyze the cell cycle and cell apoptosis.The effects of TET2 on RCC growth in vivo was analyzed using a mouse xenograft model.We found that TET2 and miR-200c were decreased in RCC tissues,and hypermethylation of miR-200c promoter was found.Overexpression of TET2 promoted miR-200c expression by reducing miR-200c promoter methylation.Additionally,overexpression of TET2 or miR-200c suppressed cell growth and metastasis.Also,knockdown of miR-200c could moderate TET2 mediated cell growth inhibition.Furthermore,we found miR-200c directly regulates Stearoyl-CoA desaturase(SCD)gene expression.Moreover,in vivo experiment results confirmed that TET2 inhibited tumor growth.In conclusion,TET2 acts as an antioncogene in RCC by regulating the miR-200c-SCD axis and providing a potential target for RCC diagnosis and treatment.
文摘目的应用hIAP-2特异性siRNA干扰肾癌GRC-1细胞hIAP-2基因的表达,探讨siRNA-hIAP-2联合姜黄素对肾癌GRC-1细胞增殖和凋亡的影响。方法体外化学合成针对hIAP-2的siRNA,通过阳离子脂质体将siRNA-hIAP-2转染肾癌GRC-1细胞,实时荧光定量PCR检测hIAP-2 m RNA表达水平,免疫印迹检测其蛋白表达水平,MTT法及流式细胞术分别测定siRNA-hIAP-2联合姜黄素处理后对肾癌GRC-1细胞增殖及凋亡的影响。结果靶向hIAP-2的siRNA可以有效抑制肾癌GRC-1细胞的hIAP-2的表达,siRNA-hIAP-2和姜黄素均可抑制GRC-1细胞增殖并诱导其一定程度的凋亡,当联合应用靶向hIAP-2的siRNA和姜黄素时,上述作用显著增强(P<0.05)。结论体外化学合成的siRNA-hIAP-2可以有效抑制肾癌GRC-1细胞的hIAP-2的表达,并能显著增强姜黄素诱导肾癌细胞GRC-1凋亡的敏感度,提示hIAP-2可作为姜黄素治疗肾癌的有效增敏靶点。
基金supported by grants from the National Natural Science Foundation of China(81972377,82025027,31301131,81972723)the Science and Technology Project of Xuzhou(KC19064)+1 种基金the Social Development Project of Jiangsu Province(BE2019644)the Natural Science Foundation of the Jiangsu Higher Education Institutions(18KJA320012)。
文摘The tumor suppressor p53 transactivates the expression of multiple genes to exert its multifaceted functions and ultimately maintains genome stability.Thus,cancer cells develop various mechanisms to diminish p53 expression and bypass the cell cycle checkpoint.In this study,we identified the gene encoding RNAbinding protein cytoplasmic polyadenylation element-binding protein 2(CPEB2)as a p53 target.In turn,CPEB2 decreases p53 messenger RNA stability and translation to fine-tune p53 level.Specifically,we showed that CPEB2 binds the cytoplasmic polyadenylation elements in the p5330-untranslated region,and the RNA recognition motif and zinc finger(ZF)domains of CPEB2 are required for this binding.Furthermore,we found that CPEB2 was upregulated in renal cancer tissues and promotes the renal cancer cell proliferation and migration.The oncogenic effect of CPEB2 is partially dependent on negative feedback regulation of p53.Overall,we identify a novel regulatory feedback loop between p53 and CPEB2 and demonstrate that CPEB2 promotes tumor progression by inactivating p53,suggesting that CPEB2 is a potential therapeutic target in human renal cancer.
