CAJ | 학술논문

目的：微小RNA（microRNA， miRNA）-10b被证实可促进多种肿瘤细胞的生长及增强其侵袭能力。文中旨在研究miRNA-10b对人低转移肺癌细胞95-C的增殖及侵袭力的影响及其作用机制。方法用脂质体法将miRNA-10b真核表达质粒瞬时转染入95-C，实验设置空白对照组、阴性对照组和miRNA-10b质粒转染组，实时定量荧光PCR（Real-Time PCR， RT-PCR，）检测各组细胞miRNA-10b的表达及KLF4mRNA的表达，细胞增殖实验检测各组细胞的增殖情况， Transwell实验检测各组细胞侵袭能力，Western blot检测各组细胞KLF4蛋白的表达。结果转染后48 h，RT-PCR检测，miRNA-10b质粒转染组miRNA-10b表达（1．61±0．12）较空白对照组（1．01±0．08）、阴性对照组（0．86±0．07）明显上调（P＜0．05），阴性对照组与空白对照组差异无统计学意义（P＞0．05）。生长曲线反映miRNA-10b质粒转染组细胞生长速度明显高于空白对照组和阴性对照组，差异有统计学意义（P＜0．05），空白对照组和阴性对照组比较，差异无统计学意义（P＞0．05）；Transwell结果显示， miRNA-10b质粒转染组[（188．0±15．1）/高倍显微镜]较空白对照组[（151．0±11．3）/高倍显微镜]、阴性对照组[（136．0±10．8）/高倍显微镜]有更多的细胞迁移到Transwell膜的另一侧（P＜0．05），空白对照组与阴性对照组差异无统计学意义（P＞0．05）；转染后48 h，miRNA-10b质粒组KLF4蛋白表达（0．86±0．06）较空白对照组（1．48±0．05）和阴性对照组（1．54±0．08）降低（P＜0．05）；各组细胞KLF4 mRNA表达差异无统计学意义（P＞0．05）。结论 miRNA-10b可能过下调KLF4蛋白的表达促进95-C细胞的增殖及侵袭能力。
목적：미소RNA（microRNA， miRNA）-10b피증실가촉진다충종류세포적생장급증강기침습능력。문중지재연구miRNA-10b대인저전이폐암세포95-C적증식급침습력적영향급기작용궤제。방법용지질체법장miRNA-10b진핵표체질립순시전염입95-C，실험설치공백대조조、음성대조조화miRNA-10b질립전염조，실시정량형광PCR（Real-Time PCR， RT-PCR，）검측각조세포miRNA-10b적표체급KLF4mRNA적표체，세포증식실험검측각조세포적증식정황， Transwell실험검측각조세포침습능력，Western blot검측각조세포KLF4단백적표체。결과전염후48 h，RT-PCR검측，miRNA-10b질립전염조miRNA-10b표체（1．61±0．12）교공백대조조（1．01±0．08）、음성대조조（0．86±0．07）명현상조（P＜0．05），음성대조조여공백대조조차이무통계학의의（P＞0．05）。생장곡선반영miRNA-10b질립전염조세포생장속도명현고우공백대조조화음성대조조，차이유통계학의의（P＜0．05），공백대조조화음성대조조비교，차이무통계학의의（P＞0．05）；Transwell결과현시， miRNA-10b질립전염조[（188．0±15．1）/고배현미경]교공백대조조[（151．0±11．3）/고배현미경]、음성대조조[（136．0±10．8）/고배현미경]유경다적세포천이도Transwell막적령일측（P＜0．05），공백대조조여음성대조조차이무통계학의의（P＞0．05）；전염후48 h，miRNA-10b질립조KLF4단백표체（0．86±0．06）교공백대조조（1．48±0．05）화음성대조조（1．54±0．08）강저（P＜0．05）；각조세포KLF4 mRNA표체차이무통계학의의（P＞0．05）。결론 miRNA-10b가능과하조KLF4단백적표체촉진95-C세포적증식급침습능력。
Objective MiRNA-10b is an important member of the MiRNA family , which has been proven that miRNA-10b can promote the growth and invasion of a variety of tumor cells .The aim of this study was to to investigate the effect and mechanism of miR-NA-10b on proliferation and invasion of low metastasis of lung cancer cell line (95-C). Methods The recombinant of miRNA-10b was transfected into 95-C by lipofectin method .The experiment set up 3 groups:blank control group , negative control group and miRNA-10b expression plasmid transfected group .MiRNA-10b expression level and KLF4mRNA expression level were detected by real-time fluores-cence quantitative PCR ( RTFQ-PCR) .The cell proliferation was detected by cell proliferation assay .The invasive ability of cell was de-tected by Transwell experiment .The expression of KLF4 protein was assessed by Western blot . Results At 48 hours after transfection, compared with blank control group (1.01 ±0.08) and negative control group (0.86 ±0.07), the miRNA-10b expression level in miRNA-10b expression plasmid transfected group (1.61 ±0.12) increased significantly (P<0.05) and there was no statistical difference between blank control group and negative control group (P>0.05).From the growth curve, the cell proliferation rate was obviously increased in miRNA-10b expression plasmid transfected group ([188.0 ±15.1]/HP) compared with the other two groups ([151.0 ±11.3]/HP), ([136.0 ±10.8]/HP) (P <0.05) and there was no statistical difference between these two groups ( P >0.05 ).Transwell result showed more cells transferred to the other side of Transwell compared with the other two groups ( P <0.05 ) and there was no statistical difference between these two groups (P >0.05).The expression of KLF4 protein decreased in miRNA-10b expression plasmid transfected group compared with the other two groups ( P<0.05).KLF4mRNA expression decreased, but the difference had no statistical significance (P>0.05). Conclusion MiRNA-10b might promote the pro-liferation and invasion of 95-C through down-regulation of KLF4 protein expression .