基础医学与临床
基礎醫學與臨床
기출의학여림상
Basic & Clinical Medicine
2015年
2期
208-212
,共5页
周荣秒%李宾%牛朝旭%王娜%黄茜%霍向然%李琰
週榮秒%李賓%牛朝旭%王娜%黃茜%霍嚮然%李琰
주영초%리빈%우조욱%왕나%황천%곽향연%리염
PLCε1基因%食管癌%RNA干扰%增殖
PLCε1基因%食管癌%RNA榦擾%增殖
PLCε1기인%식관암%RNA간우%증식
PLCε1 gene%esophageal carcinoma%RNA interference%proliferation
目的：探讨沉默PLCε1基因对食管癌Eca109细胞增殖和细胞周期的影响及其可能的机制。方法 PLCε11、PLCε12和PLCε13质粒表达载体用于沉默PLCε1，通用阴性对照质粒表达载体HK作为对照。用阳离子脂质体进行转染，筛选出干扰效果最好的质粒表达载体（ PLCε12）。实验分为Eca109组、HK组和PLCε12组。 MTT检测细胞的存活率。 FCM检测细胞周期，RT-PCR检测细胞P16、CyclinD1基因 mRNA表达。结果 HK、PLCε12质粒表达载体转染Eca109细胞后48和72 h，PLCε12组Eca109细胞的存活率分别为80.73％和75.88％，显著低于HK组（ P＜0.001）。 Eca109细胞转染后24 h，PLCε12组处于S期的细胞比例明显低于HK组（ P＜0.01），细胞周期阻滞于G0／G1期。质粒表达载体转染Eca109细胞48 h后，PLCε12组Eca109细胞P16基因mRNA表达水平明显高于HK组（ P＜0.01）。结论沉默PLCε1基因可能通过上调Eca109细胞P16基因的表达，阻止细胞周期从G1期向S期的过渡，抑制Eca109细胞的增殖活性。
目的：探討沉默PLCε1基因對食管癌Eca109細胞增殖和細胞週期的影響及其可能的機製。方法 PLCε11、PLCε12和PLCε13質粒錶達載體用于沉默PLCε1，通用陰性對照質粒錶達載體HK作為對照。用暘離子脂質體進行轉染，篩選齣榦擾效果最好的質粒錶達載體（ PLCε12）。實驗分為Eca109組、HK組和PLCε12組。 MTT檢測細胞的存活率。 FCM檢測細胞週期，RT-PCR檢測細胞P16、CyclinD1基因 mRNA錶達。結果 HK、PLCε12質粒錶達載體轉染Eca109細胞後48和72 h，PLCε12組Eca109細胞的存活率分彆為80.73％和75.88％，顯著低于HK組（ P＜0.001）。 Eca109細胞轉染後24 h，PLCε12組處于S期的細胞比例明顯低于HK組（ P＜0.01），細胞週期阻滯于G0／G1期。質粒錶達載體轉染Eca109細胞48 h後，PLCε12組Eca109細胞P16基因mRNA錶達水平明顯高于HK組（ P＜0.01）。結論沉默PLCε1基因可能通過上調Eca109細胞P16基因的錶達，阻止細胞週期從G1期嚮S期的過渡，抑製Eca109細胞的增殖活性。
목적：탐토침묵PLCε1기인대식관암Eca109세포증식화세포주기적영향급기가능적궤제。방법 PLCε11、PLCε12화PLCε13질립표체재체용우침묵PLCε1，통용음성대조질립표체재체HK작위대조。용양리자지질체진행전염，사선출간우효과최호적질립표체재체（ PLCε12）。실험분위Eca109조、HK조화PLCε12조。 MTT검측세포적존활솔。 FCM검측세포주기，RT-PCR검측세포P16、CyclinD1기인 mRNA표체。결과 HK、PLCε12질립표체재체전염Eca109세포후48화72 h，PLCε12조Eca109세포적존활솔분별위80.73％화75.88％，현저저우HK조（ P＜0.001）。 Eca109세포전염후24 h，PLCε12조처우S기적세포비례명현저우HK조（ P＜0.01），세포주기조체우G0／G1기。질립표체재체전염Eca109세포48 h후，PLCε12조Eca109세포P16기인mRNA표체수평명현고우HK조（ P＜0.01）。결론침묵PLCε1기인가능통과상조Eca109세포P16기인적표체，조지세포주기종G1기향S기적과도，억제Eca109세포적증식활성。
Objective To explore the impact of silencing PLCε1 gene on proliferation and cell cycle of esophageal carci-noma Eca109 cells.Methods Three plasmid expression vectors (PLCε11, PLCε12 and PLCε13) were constructed to si-lence PLCε1 gene.A negative control plasmid expression vector (HK) was constructed at the same time to serve as a control .The plasmid expression vectors were transfected into esophageal carcinoma Eca 109 cells by cations liposome . The plasmid expression vector with the best interference effect ( PLCε12 ) was chosen .The study included Eca 109 group , HK group and PLCε12 group .Cell viability of Eca 109 cells was evaluated by MTT assay .The cell cycles were detected by FCM .The mRNA expression of P16 and CyclinD1 gene was measured by RT-PCR.Results The cell vi-abilitys of Eca109 cells in PLCε12 group were 80.73%and 75.88%at 48 and 72 h after transfection , which were significantly lower than that of Eca 109 cells in HK group (P<0.001).The percentage of S phase Eca109 cells in PLCε12 group was lower than that of Eca 109 cells in HK group ( P <0.01 ) , the cell cycle of PLCε12 group Eca109 cells was arrested in G0/G1 phase.The P16 gene mRNA expression of PLCε12 group Eca109 cells was higher than that of HK group Eca 109 cells ( P<0.01 ) .Conclusions Silencing PLCε1 gene may up-regulate P16 gene mRNA expression and then arrest the cell cycle at G 0/G1 phase and so inhibit proliferation of Eca 109 cells.
