[1]肖正红),谢广伦),翟焕阁),等.干扰Twist表达对肾癌细胞糖酵解水平的影响[J].郑州大学学报(医学版),2019,(06):891-894.[doi:10.13705/j.issn.1671-6825.2018.07.161]
 XIAO Zhenghong),XIE Guanglun),ZHAI Huange),et al.Effects of interfering Twist expression on glycolysis metabolic level of renal carcinoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2019,(06):891-894.[doi:10.13705/j.issn.1671-6825.2018.07.161]
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干扰Twist表达对肾癌细胞糖酵解水平的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2019年06期
页码:
891-894
栏目:
应用研究
出版日期:
2019-11-20

文章信息/Info

Title:
Effects of interfering Twist expression on glycolysis metabolic level of renal carcinoma cells
作者:
肖正红1)谢广伦2)翟焕阁1)王苏芳1)
1)南阳南石医院肿瘤内科 河南南阳 473065 2)郑州大学附属肿瘤医院疼痛科 郑州450008
Author(s):
XIAO Zhenghong1)XIE Guanglun2)ZHAI Huange1)WANG Sufang1)
1)Department of Oncology,Nanyang Nanshi Hospital,Nanyang,Henan 473065 2)Department of Pain,Cancer Hospital Affiliated to Zhengzhou University,Zhengzhou 450008
关键词:
肾癌 Twist 糖酵解
Keywords:
renal carcinoma Twist glycolysis
分类号:
R737.11
DOI:
10.13705/j.issn.1671-6825.2018.07.161
摘要:
目的:研究干扰Twist表达对肾癌细胞糖酵解水平的影响。方法:用重组Twist shRNA慢病毒(Twist shRNA组)和阴性对照慢病毒(shRNA-NC组)感染肾癌细胞A498,用Real-time PCR和Western blot法测定Twist的表达,MTT法测定细胞存活率,平板克隆实验检测克隆形成能力,Western blot法检测细胞中己糖激酶2(HK2)和丙酮酸激酶M2亚型(PKM2)蛋白表达水平,用乳酸脱氢酶比色法检测乳酸分泌水平,用葡萄糖氧化酶法检测葡萄糖消耗量,用比色法检测细胞中ATP含量。以未感染慢病毒的细胞为空白对照。结果:与空白对照组和shRNA-NC组比较,Twist shRNA组肾癌细胞存活率和克隆形成率,细胞中HK2和PKM2蛋白表达水平,乳酸分泌水平,葡萄糖消耗量,以及细胞中ATP含量均降低(P<0.05)。结论:下调Twist可抑制肾癌细胞糖酵解关键酶的表达,降低细胞能量代谢水平,抑制细胞生长。
Abstract:
Aim:To study the effects of interfering Twist expression on glycolysis metabolic level of renal carcinoma cells.Methods: Renal carcinoma A498 cells were infected with recombinant lentivirus loaded Twist shRNA(Twist shRNA group)or negative control recombinant lentivirus(shRNA-NC group), the expressions of Twist mRNA and protein were detected by real-time PCR and Western blot, cell survival was detected by MTT method, cell clone formation ability was detected by plate cloning assay,and Western blot was used to detect the levels of HK2 and PKM2 proteins; lactic acid dehydrogenase colorimetry was used to detect the level of lactic acid secretion, glucose consumption was detected by glucose oxidase, and ATP content was detected by colorimetry. The cells without infection were the blank control.Results:Compared with shRNA-NC group and blank control group,the survival, cell clone formation ability, the levels of HK2 and PKM2 protein expression, the content of lactic acid secretion, glucose consumption, and the level of ATP in cells were decreased in Twist shRNA group(P<0.05).Conclusion:Down-regulation of Twist could inhibit the expressions of key glycolytic enzymes in renal carcinoma cells, reduce the level of cell energy metabolism, and inhibit cell growth.

参考文献/References:

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备注/Memo

备注/Memo:
【基金项目】河南省科技攻关项目(172102310688) 【作者简介】肖正红,女,1974年7月生,本科,副主任医师,研究方向:肿瘤治疗,E-mail:utub621627@163.com
更新日期/Last Update: 2019-11-20