天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (9): 818-823.doi: 10.11958/20193951

• 细胞与分子生物学 • 上一篇    下一篇

SIRT5对高糖环境下结肠癌细胞增殖、凋亡、迁移的影响及机制探讨

贺帅1，岳淑芬2，周蕾3，齐琦4   

  1. 1包头医学院病理学教研室，包头医学院第一附属医院病理科（邮编014060）；2包头医学院组织学与胚胎学教研室；3包头医学院病原生物与免疫学实验室；4包头医学院2014级临床医学本科
  • 收稿日期:2019-12-30 修回日期:2020-05-26 出版日期:2020-09-15 发布日期:2020-09-15
  • 作者简介:贺帅（1980），女，硕士，副教授，主要从事肿瘤发病机制的研究
  • 基金资助:
    内蒙古自然科学基金项目［2017MS（LH）0813］；包头市卫生基金项目（Wsjj2017076）

Effects and mechanism of SIRT5 on the proliferation, apoptosis and migration of colon cancer cells in high glucose environment#br#

 HE Shuai1, YUE Shu-fen2, ZHOU Lei3, QI qi4   

  1. 1 Department of Pathology, Baotou Medical College, Baotou 014060, China; 2 Department of Pathology, the First Affiliated Hospital of Baotou Medical College; 3 Laboratory of Pathogen and Immunology, 4 Grade 2014 Undergraduate of Clinical Medicine, Baotou Medical College
  • Received:2019-12-30 Revised:2020-05-26 Published:2020-09-15 Online:2020-09-15

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摘要： 摘要:目的　探讨Sirtuin5（SIRT5）对高糖环境下Lovo细胞增殖、凋亡及迁移的影响及可能机制。方法　高糖培养基（葡萄糖含量4 500 mg/L）及普通培养基（葡萄糖含量2 000 mg/L）培养SIRT5高表达的人结肠癌Lovo细胞，进行慢病毒转染实验，高糖培养基及普通培养基各分为空白对照组（MOCK组，无干预）、阴性对照组（NC组，对照RNAi慢病毒转染）和SIRT5-RNAi组（KD组，SIRT5-RNAi慢病毒转染），用MTT法检测各组细胞1~5 d增殖能力变化，AnnexinⅤ检测各组细胞凋亡率，划痕实验法检测各组细胞迁移率；应用实时荧光定量PCR（qPCR）检测各组细胞SIRT5、哺乳动物雷帕霉素靶蛋白（mTOR）、缺氧诱导因子-1α（HIF-1α）、M2型丙酮酸激酶（PKM2）及己糖激酶-2（HK2）mRNA的表达变化，Western blot检测各组上述指标的蛋白表达水平。结果　高糖培养基中，KD组与NC组、MOCK组比较，3~5 d时增殖水平降低，凋亡率增加，而24 h迁移率降低（P＜0.05），且SIRT5、mTOR、HIF-1α、PKM2及HK2 mRNA及蛋白的相对表达水平均明显降低（P＜0.05）。普通培养基中，KD组与NC及MOCK组比较，1~4 d时增殖水平差异无统计学意义，5 d时增殖水平降低（P＜0.05），24 h凋亡率差异无统计学意义。结论　高糖环境下，SIRT5可能通过影响糖代谢关键酶HK2及mTOR/HIF-1α/PKM2信号通路调控糖酵解，从而促进结肠癌的发生、发展。

关键词: 结肠肿瘤；细胞系, 肿瘤；抗衰老酶；缺氧诱导因子1, α亚基；丙酮酸激酶；己糖激酶；高糖；哺乳动物雷帕霉素靶蛋白； sirtuin 5

Abstract: Abstract: Objective　To explore the effect of Sirtuin5 (SIRT5) on the proliferation, apoptosis and migration of Lovo cells in high glucose environment and its possible mechanism. Methods　Lovo cells with high expression of SIRT5 were cultured in vitro with high glucose medium (4 500 mg/L) and ordinary medium (2 000 mg/L). Lentivirus transfection experiment was performed. SIRT5-RNAi transfected with SIRT5-RNAi lentivirus was used as KD group, blank control group with no intervention was used as MOCK group, and negative control group transfected with control RNAi lentivirus was used as NC group. The cell proliferation was observed by MTT assay, apoptosis rate was determined by flow cytometry, and cell migration rate was measured by scratch test. The expressions of mammalian target of rapamycin (mTOR), hypoxia inducible factor-1α (HIF-1α), M2 pyruvate kinase (PKM2) and hexokinase2 (HK2) were detected by real-time PCR, and their protein expressions were detected by Western blot assay. Results　Compared with NC and MOCK, the proliferation decreased at the 3-5 day following SIRT5 silence, the cell apoptosis rate increased and the cell migration rate decreased in KD with high glucose medium (P＜0.01). The proliferation decreased at the 5-day following SIRT5 silence (P＜0.05), but no significant difference in the cell apoptosis rate between KD and NC groups at the 1-4 day. The expressions of SIRT5, mTOR, HIF-1 α, PKM2, HK2 mRNA and proteins were significantly lower in KD group than those in NC and MOCK groups with high glucose medium (P＜0.05). Conclusion　SIRT5 promotes the development of colon cancer in high glucose environment, which may be related with the regulating glycolysis by affecting the key glucose metabolism enzymes HK2 and mTOR/HIF-1α/PKM2 signal pathway. 

Key words: colonic neoplasms, cell line,tumor;sirtuins, hypoxia-inducible factor 1,alpha subunit, pyruvate kinase, hexokinase, high glucose, mammalian target of rapamycin, sirtuin 5

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