摘要
目的探讨上调微小RNA-141模拟物（miR-141mimics）表达对肝癌细胞（HepG2）增殖、凋亡及细胞周期的影响。方法采用实时荧光定量PCR（qPCR）检测正常肝上皮细胞（L02）和肝癌细胞（HepG2）中miR-141的差异表达。以合成的miR-141模拟物转染HepG2细胞作为实验组（miR-141组），以空白对照（CON组）和阴性对照（miR—NC组）作为对照组。采用qPCR检测各组细胞中miR-141表达情况，CCK-8法检测细胞的增殖情况，流式细胞仪检测细胞凋亡和细胞周期。结果qPCR结果显示，HepG2中miR-141相对表达（0．64±0．13）明显低于L02（1．00±0．18），差异有统计学意义（P〈0．05）。转染后，miR-141组细胞中miR-141相对表达（3．33±0．66）明显高于CON组（1．00±0．17，P〈0．05）和miR—NC组（1．08±0．14，P〈0．05）。CCK8结果显示，miR-141组在转染后48、72、96h相对吸光度值分别为（0．67±0．07）、（1．17±0．05）、（1．36±0．03），均明显低于CON组[（0．81±0．02）、（1．42±0．03）、（1．73±0．05）]和miR—NC组[（0．78±0．01）、（1．38±0．02）、（1．69±0．01），均P〈0．05]。细胞凋亡检测结果显示，miR-141组细胞凋亡率[（11．81±0．23）％]明显高于CON组[（4．18±0．18）％]和miR—NC组[（4．04±0．08）％]，差异有统计学意义（P〈0．05）。细胞周期检测结果显示，miR-141组S期细胞[（19．89±2．78）％]明显少于CON组[（31．87±1．00）％]和miR—NC组[（30．49±1．73）％]，差异均有统计学意义（均P〈0．05）；而miR-141组GO／G1期细胞[（74．74±2．03）％]明显多于CON组[（60．85±1．69）％]和miR．NC组[（60．93±1．95）％]，差异均有统计学意义（均P〈0．05）。结论HepG2细胞中miR-141低表达。上调miR-141表达能明显抑制HepG2细胞增殖，促进细胞凋亡及改变细胞周期的分布。
Objective To investigate the effect of iniR-141 up-regulation through miR-141 mimic transfection on proliferation, apoptosis and cell cycle of hepatocellular carcinoma （HCC） cells. Methods Real-time polymerase chain reaction （qPCR） was employed to detect differential expression of miR-141 in normal hepatic epithelial cells （L02） and hepatocellular carcinoma cells （HepG2）. Furthermore, miR-141 mimics were used to transfect into HepG2 to up-regulate the expression of miR-141 （experimental group）. Blank group （CON） and negative control group （miR-NC） were used as control group, qPCR was used to detect expression of miR-141. CCK8 assay was used to detect the proliferation of HepG2 cells. Flow cytome- try was used to detect the apoptosis and cell cycle of HepG2 cells. Results The results of qPCR showed that miR-141 was significantly down-regulated in HepG2 cells （0. 64±0. 13 ） compared to L02 cells （ 1.00±0. 18 ） , and the difference was significant （ P 〈 0.05 ）. Relative expression of miR-141 was significantly increased after transfection （3.33 ± 0.66）, compared to CON group （ 1.00 ±0. 17, P 〈 0.05 ） and miR-NC group （ 1.08±0.14 ,P 〈 0.05 ）. CCK8 assay showed that the absorbance values of HepG2 of miR- 141 group at 48, 72, 96 h （0.67 ±0.07, 1.17 ± 0.05, 1.36 ± 0.03 ） were all decreased significantly compared with those in CON group （0.81±0.02 ; 1.42 ± 0.03 ; 1.73 ±0. 05, all P 〈 0. 05 ） and miR-NC group （0. 78± 0. 01 ; 1.38 ± 0.02 ; 1.69± 0. 01, all P 〈 0.05 ）. The results showed that miR-141 group [ （11.81 ± 0.23 ）% ] had significantly increased apoptosis rate compared to CON group [ （4. 18 ± 0. 18） % ] and miR-NC group [ （ 4.04 ± 0. 08 ） % ], and the difference was statistically significant （ P 〈0. 05）. miR-141 group had decreased percentage of S phase cells [ （ 19. 89 ± 2. 78） % ] compared to CON group [ （31.87 ± 1.00） %, P 〈 0. 05 ]and miR-NC group [ （ 30. 49 ± 1.73 ） %, P 〈 0. 05 ], whi
出处
《中华肝胆外科杂志》
CSCD
北大核心
2017年第6期401-405,共5页
Chinese Journal of Hepatobiliary Surgery
