目的:探讨过表达MARCH2对肝细胞癌HUH7细胞自噬水平和生长的影响及其机制。方法:MTS和EDU检测MARCH2过表达的肝细胞癌HUH7细胞的生长情况。流式细胞术检测MARCH2过表达的肝细胞癌HUH7细胞凋亡、细胞周期的情况。Western blot和激光共聚...目的:探讨过表达MARCH2对肝细胞癌HUH7细胞自噬水平和生长的影响及其机制。方法:MTS和EDU检测MARCH2过表达的肝细胞癌HUH7细胞的生长情况。流式细胞术检测MARCH2过表达的肝细胞癌HUH7细胞凋亡、细胞周期的情况。Western blot和激光共聚焦法检测MARCH2过表达的肝细胞癌HUH7细胞自噬水平。结果:MTS和EDU检测表明,MARCH2的过表达促进了肝细胞癌HUH7的生长。流式细胞术检测表明,MARCH2过表达对肝细胞癌HUH7细胞凋亡和细胞周期没有明显影响。进一步的实验表明,MARCH2过表达损害了肝细胞癌HUH7的自噬。用常见的自噬诱导剂雷帕霉素挽救自噬可以使肝细胞癌HUH7的增殖率恢复到控制水平。结论:我们的数据表明,MARCH2通过抑制自噬在肿瘤发展过程中起着至关重要的作用。因此,MARCH2是一种有前景的肿瘤治疗新靶点。Aims: To investigate the effects and mechanisms of overexpression of MARCH2 on autophagy levels and growth of hepatocellular carcinoma HUH7 cells. Methods: MTS and EDU were used to detect the growth of MARCH2 overexpressing hepatocellular carcinoma HUH7 cells. Flow cytometry was used to detect apoptosis and cell cycle in MARCH2 overexpressing hepatocellular carcinoma HUH7 cells. Western blot and laser confocal microscopy were used to detect the autophagy levels in MARCH2 overexpressing HUH7 hepatocellular carcinoma cells. Results: In this study, MTS and EDU assays showed that the overexpression of MARCH2 promoted the growth of hepatocellular carcinoma HUH7 cells. Flow cytometry detection illustrated that MARCH2 overexpression had no obvious effect on the hepatocellular carcinoma HUH7 cell apoptosis and cell cycle. Further experiments displayed that MARCH2 overexpression impaired the autophagy of hepatocellular carcinoma HUH7 cells. Rescued autophagy with the common autophagy inducer rapamycin can bring the proliferation rate of hepatocellular carcinoma HUH7 cells back to the control level. Conclusions: Our data indicates that MARCH2 plays an essential role in the progression of tumor development through inhibiting autophagy. Therefore, MARCH2 is an attractive novel target for tumor therapy.展开更多
文摘目的:探讨过表达MARCH2对肝细胞癌HUH7细胞自噬水平和生长的影响及其机制。方法:MTS和EDU检测MARCH2过表达的肝细胞癌HUH7细胞的生长情况。流式细胞术检测MARCH2过表达的肝细胞癌HUH7细胞凋亡、细胞周期的情况。Western blot和激光共聚焦法检测MARCH2过表达的肝细胞癌HUH7细胞自噬水平。结果:MTS和EDU检测表明,MARCH2的过表达促进了肝细胞癌HUH7的生长。流式细胞术检测表明,MARCH2过表达对肝细胞癌HUH7细胞凋亡和细胞周期没有明显影响。进一步的实验表明,MARCH2过表达损害了肝细胞癌HUH7的自噬。用常见的自噬诱导剂雷帕霉素挽救自噬可以使肝细胞癌HUH7的增殖率恢复到控制水平。结论:我们的数据表明,MARCH2通过抑制自噬在肿瘤发展过程中起着至关重要的作用。因此,MARCH2是一种有前景的肿瘤治疗新靶点。Aims: To investigate the effects and mechanisms of overexpression of MARCH2 on autophagy levels and growth of hepatocellular carcinoma HUH7 cells. Methods: MTS and EDU were used to detect the growth of MARCH2 overexpressing hepatocellular carcinoma HUH7 cells. Flow cytometry was used to detect apoptosis and cell cycle in MARCH2 overexpressing hepatocellular carcinoma HUH7 cells. Western blot and laser confocal microscopy were used to detect the autophagy levels in MARCH2 overexpressing HUH7 hepatocellular carcinoma cells. Results: In this study, MTS and EDU assays showed that the overexpression of MARCH2 promoted the growth of hepatocellular carcinoma HUH7 cells. Flow cytometry detection illustrated that MARCH2 overexpression had no obvious effect on the hepatocellular carcinoma HUH7 cell apoptosis and cell cycle. Further experiments displayed that MARCH2 overexpression impaired the autophagy of hepatocellular carcinoma HUH7 cells. Rescued autophagy with the common autophagy inducer rapamycin can bring the proliferation rate of hepatocellular carcinoma HUH7 cells back to the control level. Conclusions: Our data indicates that MARCH2 plays an essential role in the progression of tumor development through inhibiting autophagy. Therefore, MARCH2 is an attractive novel target for tumor therapy.
