浙江中西医结合杂志
浙江中西醫結閤雜誌
절강중서의결합잡지
ZHEJIANG JOURNAL OF INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE
2015年
4期
341-344
,共4页
肝肿瘤%Huh7%Mcl-1%细胞增殖%细胞凋亡%miR-29b
肝腫瘤%Huh7%Mcl-1%細胞增殖%細胞凋亡%miR-29b
간종류%Huh7%Mcl-1%세포증식%세포조망%miR-29b
Hepatocellular carcinoma%Huh7%Mcl-1%Cell proliferation%Apoptosis%miR-29b
目的：探讨miR-29b（microRNA-29b）的作用靶点及其对肝肿瘤细胞系Huh7细胞增殖及凋亡的干预作用。方法荧光定量PCR检测正常肝细胞系LO2及肝癌细胞系Huh7的miR-29b和Mcl-1表达水平。通过生物信息学分析预测Mcl-1基因是否受microRNA调控。将miR-29b模拟物通过Lipofectamine 2000顺时转染Huh7细胞，检测miR-29b对Mcl-1表达水平的影响。MTT法检测miR-29b模拟物转染对Huh7细胞增殖的影响。Annexin V/PI染色法检测miR-29b对Huh7细胞凋亡的影响。结果肝肿瘤细胞相比于正常肝细胞高表达Mcl-1（3.42依0.15比1.00依0.04，P<0.05）低表达miR-29b（0.43依0.04比1.00依0.06，P<0.05），在肝癌细胞中转染miR-29b可使Mcl-1的表达水平下降（0.37依0.03比1.03依0.07，P<0.05），转染miR-29b可抑制Huh7细胞的增殖（0.85依0.11比1.68依0.17，P<0.05）并诱导其发生凋亡[（28.4依2.10）%比（3.6依0.06）%，P<0.05]。结论 MiR-29b下调Huh7细胞Mcl-1蛋白的表达并诱导其发生凋亡。
目的：探討miR-29b（microRNA-29b）的作用靶點及其對肝腫瘤細胞繫Huh7細胞增殖及凋亡的榦預作用。方法熒光定量PCR檢測正常肝細胞繫LO2及肝癌細胞繫Huh7的miR-29b和Mcl-1錶達水平。通過生物信息學分析預測Mcl-1基因是否受microRNA調控。將miR-29b模擬物通過Lipofectamine 2000順時轉染Huh7細胞，檢測miR-29b對Mcl-1錶達水平的影響。MTT法檢測miR-29b模擬物轉染對Huh7細胞增殖的影響。Annexin V/PI染色法檢測miR-29b對Huh7細胞凋亡的影響。結果肝腫瘤細胞相比于正常肝細胞高錶達Mcl-1（3.42依0.15比1.00依0.04，P<0.05）低錶達miR-29b（0.43依0.04比1.00依0.06，P<0.05），在肝癌細胞中轉染miR-29b可使Mcl-1的錶達水平下降（0.37依0.03比1.03依0.07，P<0.05），轉染miR-29b可抑製Huh7細胞的增殖（0.85依0.11比1.68依0.17，P<0.05）併誘導其髮生凋亡[（28.4依2.10）%比（3.6依0.06）%，P<0.05]。結論 MiR-29b下調Huh7細胞Mcl-1蛋白的錶達併誘導其髮生凋亡。
목적：탐토miR-29b（microRNA-29b）적작용파점급기대간종류세포계Huh7세포증식급조망적간예작용。방법형광정량PCR검측정상간세포계LO2급간암세포계Huh7적miR-29b화Mcl-1표체수평。통과생물신식학분석예측Mcl-1기인시부수microRNA조공。장miR-29b모의물통과Lipofectamine 2000순시전염Huh7세포，검측miR-29b대Mcl-1표체수평적영향。MTT법검측miR-29b모의물전염대Huh7세포증식적영향。Annexin V/PI염색법검측miR-29b대Huh7세포조망적영향。결과간종류세포상비우정상간세포고표체Mcl-1（3.42의0.15비1.00의0.04，P<0.05）저표체miR-29b（0.43의0.04비1.00의0.06，P<0.05），재간암세포중전염miR-29b가사Mcl-1적표체수평하강（0.37의0.03비1.03의0.07，P<0.05），전염miR-29b가억제Huh7세포적증식（0.85의0.11비1.68의0.17，P<0.05）병유도기발생조망[（28.4의2.10）%비（3.6의0.06）%，P<0.05]。결론 MiR-29b하조Huh7세포Mcl-1단백적표체병유도기발생조망。
Objective To investigate the therapeutic targets of miR-29b and its effect on proliferation and apoptosis of hepatocarcinoma cell line Huh7. Methods The expression of Mcl-1 and miR-29b in normal liver cell line LO2 and hepatocellular carcinoma cell line Huh7 was detected by qPCR. The putative binding sites of Mcl-1 gene was analyzed by using bioinformatic method. MiR-29b was transfected into Huh7 cells by Lipofectamine 2000, then the expression of Mcl-1 was detected. The cell growth was assessed by MTT after Huh7 cells transfected with miR-29b mimics. The apoptosis of cells was measured by Annexin V/PI staining. Results Compared with normal liver cells, Huh7 cells had higher expression of Mcl-1 (3.42±0.15 vs 1.00±0.04, P<0.05) and lower expression of miR-29b(0.43±0.04 vs 1.00±0.06, P<0.05). Transfection of miR-29b mimics suppressed Mcl-1 expression in Huh7 cells (0.37±0.03 vs 1.03±0.07, P<0.05) and inhibited the proliferation of Huh7 cells (0.85±0.11 vs 1.68±0.17, P<0.05) while inducing apoptosis in Huh7 cells [(28.4±2.1)% vs (3.6±0.06)%, P<0.05]. Conclusion MiR-29b down-regulates the expression of Mcl-1 protein inducing apoaptosis in Huh7 cells.
