3-羟基丁酸对HeLa癌细胞生长与凋亡的影响
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摘要
研究3-羟基丁酸对HeLa癌细胞生长与凋亡的影响.采用四唑盐(MTT)比色法测定细胞活力;AO/EB荧光染色观察细胞凋亡与坏死的形态学变化;单细胞凝胶电泳检测细胞DNA的损伤;流式细胞仪检测细胞凋亡;蛋白质免疫印迹法(Western Blot)检测凋亡蛋白的表达.结果表明:3-羟基丁酸能够抑制HeLa癌细胞增殖;使HeLa癌细胞DNA受到损伤;通过caspase依赖性细胞凋亡途径诱导HeLa癌细胞凋亡;在HeLa癌细胞与人脐带正常细胞共培养条件下,3-羟基丁酸诱导HeLa癌细胞先于人脐带正常细胞凋亡.实验为肿瘤的生酮饮食疗法和酮体疗法提供了理论依据.
To study the effects of 3-hydroxybutyrate on growth and apoptosis of HeLa cells, cell viability was detected by MTT. The morphological alterations of apoptosis or necrosis in HeLa cells were stained with AO/EB. HeLa cell DNA damage was analyzed by single cell gel electrophoresis(SCGE) test and the cell apoptosis was analyzed by flow cytometry. The expression of apoptotic protein was determined by Western Blot. The results showed that 3-hydroxybutyrate could inhibit the proliferation and damage the DNA of HeLa cells. The 3-hydroxybutyrate induced HeLa cancer cell apoptosis through caspase-dependent cell apoptosis pathway. In the co-culture condition, the 3-hydroxybutyrate induced HeLa cells to apoptosis before human umbilical cord cells. The results would provide a theoretical basis for the treatment of tumors with ketogenic diet and 3-hydroxybutyrate.
To study the effects of 3-hydroxybutyrate on growth and apoptosis of HeLa cells, cell viability was detected by MTT. The morphological alterations of apoptosis or necrosis in HeLa cells were stained with AO/EB. HeLa cell DNA damage was analyzed by single cell gel electrophoresis(SCGE) test and the cell apoptosis was analyzed by flow cytometry. The expression of apoptotic protein was determined by Western Blot. The results showed that 3-hydroxybutyrate could inhibit the proliferation and damage the DNA of HeLa cells. The 3-hydroxybutyrate induced HeLa cancer cell apoptosis through caspase-dependent cell apoptosis pathway. In the co-culture condition, the 3-hydroxybutyrate induced HeLa cells to apoptosis before human umbilical cord cells. The results would provide a theoretical basis for the treatment of tumors with ketogenic diet and 3-hydroxybutyrate.
引文
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[3]郝光伟,王海玉,何德明,等.生酮饮食对人结肠癌裸鼠皮下移植瘤生长的影响[J].中国肿瘤临床,2014,4l(18):1154-1157.
[4]DUREGGER A D,RAMONER R,PANTE J,et al.Differential metabolic effects of medium-chain triglycerides and omega-3 fatty acids in benign and malignant prostate cells-evidence for a ketogenic diet as adjuvant therapy for prostate cancer[J].Eur Urol Suppl,2014,13(1):e305.
[5]HANS V,RAINER W,NADJA D,et al.Growth of human gastric cancer cells in nude mice is delayed by a ketogenic diet supplemented with omega-3 fatty acids and medium-chain triglycerides[J].BMC Cancer,2008,8(1):1-12.
[6]RUSKIN D N,KAWAMURA M,MASINO S A.Reduced pain and inflammation in juvenile and adult rats fed a ketogenic diet[J].Plos One,2009,4(12):e8349.
[7]MASINO S A,GEIGER J D.Are purines mediators of the anticonvulsant/neuroprotective effects of ketogenic diets?[J].Trends Neurosci,2008,31(6):273-278.
[8]STAFFORD P,ABDELWAHAB M G,KIM D Y,et al.The ketogenic diet reverses gene expression patterns and reduces reactive oxygen species levels when used as an adjuvant therapy for glioma[J].Nutr Metab,2010,7(1):1-11.
[9]BRYAN A G,SUDERSHAN B K,BRANDT E K,et al.Ketogenic diets enhance oxidative stress and radio-chemo-therapy responses in lung cancer xenografts[J].Clin Cancer Res,2013,19(14):3905-3913.
[10]SHUKLA S K,GEBREGIWORGIS T,PUROHIT V,et al.Metabolic reprogramming induced by ketone bodies diminishes pancreatic cancer cachexia[J].Cancer Metab,2014,2(1):18-23.
[11]KOCSIS K,FRANK R,SZABO J,et al.Acetyl-l-carnitine restores synaptic transmission and enhances the inducibility of stable LTP after oxygen-glucose deprivation[J].Neuroscience,2016,332(22):203-211.
[12]PEARSONSTUTTARD J,ZHOU B,KONTIS V,et al.Worldwide burden of cancer attributable to diabetes and high body-mass index:A comparative risk assessment[J].Lancet Diabetes Endo,2017,6(2):95-104.
[13]高海涛,邵邻相,孙桂菊.饮食限制对学习记忆和抗肿瘤作用的研究进展[J].浙江师范大学学报(自然科学版),2014,37(1):99-107.
[14]陈佳.碳源饥饿对HeLa细胞凋亡的影响[D].金华:浙江师范大学,2016.
[15]苏佳佳,曾杰,邵邻相.碳源饥饿对癌细胞和正常细胞增殖的影响[J].浙江师范大学学报(自然科学版),2017,40(3):331-335.
[16]曾杰,陈佳,邵邻相,等.葡萄糖饥饿对HeLa细胞形态与结构的影响[J].浙江师范大学学报(自然科学版),2018,41(1):92-96.
[17]张璐敏,吕学维,邵邻相,等.野艾蒿挥发油诱导HeLa细胞凋亡与坏死[J].中药材,2013(12):1988-1992.
[18]陈佳,邵邻相,麻艳芳,等.佛手挥发油诱导HeLa细胞凋亡与坏死[J].浙江师范大学学报(自然科学版),2016,39(2):215-218.
[19]SEYFRIED T N,KIEBISH M,MUKHERJEE P,et al.Targeting energy metabolism in brain cancer with calorically restricted ketogenic diets[J].Epilepsia,2008,49(s8):114-116.
[20]WALLACE D C,FAN W.The pathophysiology of mitochondrial disease as modeled in the mouse[J].Gene Dev,2009,23(15):1714-1736.