中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2013年
7期
465-469
,共5页
蒯小玲%宋梦娇%俞智华%张健锋%陈晓奇%张弘%毛振彪
蒯小玲%宋夢嬌%俞智華%張健鋒%陳曉奇%張弘%毛振彪
괴소령%송몽교%유지화%장건봉%진효기%장홍%모진표
胃肿瘤%细胞增殖%细胞运动%微RNAs
胃腫瘤%細胞增殖%細胞運動%微RNAs
위종류%세포증식%세포운동%미RNAs
Stomach neoplasms%Cell proliferation%Cell movement%MicroRNAs
目的 探讨微小核糖核酸-32(miRNA-32)转染对胃癌细胞生物学行为的影响及其作用机制.方法 应用脂质体法分别将miRNA-32类似物、miRNA-32抑制物和空质粒瞬时转染到胃癌细胞SGC-7901,分为miRNA-32类似物转染组、miRNA-32抑制物转染组、空质粒转染组、未转染组.荧光显微镜观察绿色荧光蛋白的表达,实时荧光定量PCR法检测miRNA-32 mRNA的表达,CCK-8法检测细胞增殖能力,划痕实验、Transwell小室检测各组细胞体外迁移能力.统计学处理采用单因素方差分析.结果 与未转染组、空质粒转染组相比,miRNA-32类似物转染组mRNA表达(相对定量值=2.327)明显上调,miRNA-32抑制物转染组mRNA表达(相对定量值=0.402)明显下调,差异有统计学意义(F=11.238,P＜0.05).miRNA-32类似物转染组24 h划痕宽度为(61.39±2.21) μm,miRNA-32抑制物转染组为(29.97±0.66) μm,miRNA-32抑制物转染组细胞迁移距离较miRNA-32类似物转染组远,差异有统计学意义(F=9.371,P＜0.05).转染48 h后,miRNA-32类似物转染组穿膜细胞数较未转染组明显减少,分别为(16.93±4.63)个和(93.93±7.09)个,差异有统计学意义(F=6.853,P＜0.05).转染后48、72 h,miRNA-32类似物转染组细胞生长抑制率分别为(43.474±18.636)％和(45.050±23.764)％,细胞出现明显生长抑制(F=7.986、8.635,P=0.028、0.032).结论 上调miRNA-32表达可明显抑制人胃癌细胞SGC-7901的生长,抑制其迁移能力.
目的 探討微小覈糖覈痠-32(miRNA-32)轉染對胃癌細胞生物學行為的影響及其作用機製.方法 應用脂質體法分彆將miRNA-32類似物、miRNA-32抑製物和空質粒瞬時轉染到胃癌細胞SGC-7901,分為miRNA-32類似物轉染組、miRNA-32抑製物轉染組、空質粒轉染組、未轉染組.熒光顯微鏡觀察綠色熒光蛋白的錶達,實時熒光定量PCR法檢測miRNA-32 mRNA的錶達,CCK-8法檢測細胞增殖能力,劃痕實驗、Transwell小室檢測各組細胞體外遷移能力.統計學處理採用單因素方差分析.結果 與未轉染組、空質粒轉染組相比,miRNA-32類似物轉染組mRNA錶達(相對定量值=2.327)明顯上調,miRNA-32抑製物轉染組mRNA錶達(相對定量值=0.402)明顯下調,差異有統計學意義(F=11.238,P＜0.05).miRNA-32類似物轉染組24 h劃痕寬度為(61.39±2.21) μm,miRNA-32抑製物轉染組為(29.97±0.66) μm,miRNA-32抑製物轉染組細胞遷移距離較miRNA-32類似物轉染組遠,差異有統計學意義(F=9.371,P＜0.05).轉染48 h後,miRNA-32類似物轉染組穿膜細胞數較未轉染組明顯減少,分彆為(16.93±4.63)箇和(93.93±7.09)箇,差異有統計學意義(F=6.853,P＜0.05).轉染後48、72 h,miRNA-32類似物轉染組細胞生長抑製率分彆為(43.474±18.636)％和(45.050±23.764)％,細胞齣現明顯生長抑製(F=7.986、8.635,P=0.028、0.032).結論 上調miRNA-32錶達可明顯抑製人胃癌細胞SGC-7901的生長,抑製其遷移能力.
목적 탐토미소핵당핵산-32(miRNA-32)전염대위암세포생물학행위적영향급기작용궤제.방법 응용지질체법분별장miRNA-32유사물、miRNA-32억제물화공질립순시전염도위암세포SGC-7901,분위miRNA-32유사물전염조、miRNA-32억제물전염조、공질립전염조、미전염조.형광현미경관찰록색형광단백적표체,실시형광정량PCR법검측miRNA-32 mRNA적표체,CCK-8법검측세포증식능력,화흔실험、Transwell소실검측각조세포체외천이능력.통계학처리채용단인소방차분석.결과 여미전염조、공질립전염조상비,miRNA-32유사물전염조mRNA표체(상대정량치=2.327)명현상조,miRNA-32억제물전염조mRNA표체(상대정량치=0.402)명현하조,차이유통계학의의(F=11.238,P＜0.05).miRNA-32유사물전염조24 h화흔관도위(61.39±2.21) μm,miRNA-32억제물전염조위(29.97±0.66) μm,miRNA-32억제물전염조세포천이거리교miRNA-32유사물전염조원,차이유통계학의의(F=9.371,P＜0.05).전염48 h후,miRNA-32유사물전염조천막세포수교미전염조명현감소,분별위(16.93±4.63)개화(93.93±7.09)개,차이유통계학의의(F=6.853,P＜0.05).전염후48、72 h,miRNA-32유사물전염조세포생장억제솔분별위(43.474±18.636)％화(45.050±23.764)％,세포출현명현생장억제(F=7.986、8.635,P=0.028、0.032).결론 상조miRNA-32표체가명현억제인위암세포SGC-7901적생장,억제기천이능력.
Objective To explore the effect of microRNA-32 (miRNA-32)on the biological behaviors of gastric cancer cell and its mechanism.Methods Gastric cancer cell line SGC-7901 cells were transiently transfected with miRNA-32 analogue,miRNA-32 inhibitor and empty plasmid vectors by lipofectamine and divided into analogue transfection group,inhibitor transfection group,empty plasmid transfection group and non-transfection group.The expression of green fluorescent protein was observed under fluorescent microscopy.The expression of miRNA-32 at mRNA level was detected by quantificational real-time polymerase chain reaction.The cell proliferation was evaluated by CCK-8 assay.The cell migration ability was measured by scratch test and Transwell chamber assays.The data were analyzed by one-way ANOVA.Results Compared with empty plasmid transfection group and non-transfection group,the expression of miRNA-32 mRNA of miRNA-32 analogue transfection group (relative quantitative value:2.327) was significantly up-regulated and that of miRNA-32 inhibitor transfection group (relative quantitative value:0.402) was significantly down regulated (F=11.238,P＜0.05).The width of scratch of miRNA-32 analogue transfection group was (61.39± 2.21) μm at 24 hours; miRNA-32 inhibitor transfection group was (29.97±0.66) μm.The migration distance of inhibitor transfection group was far than that of analogue transfection group (F=9.371,P＜0.05).After transfection for 48 hours,the cell number of migrated cells of analogue transfection group was significantly less than that of non transfection group,which was 16.93±4.63 and 93.93± 7.09,respectively (F=6.853,P＜0.05).After transfection for 48 hours and 72 hours,the cell growth inhibiting rate of miRNA 32 analogue transfection group was (43.474 ± 18.636)％ and (45.050±23.764)％,respectively,the cell growth was significantly inhibited (F=7.986 and 8.635,P=0.028 and 0.032).Conclusion The cell growth and migration ability of human gastric cancer cell line SGC-7901 are obviously inhibited through upregulating the expression of miRNA-32.
