肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2014年
11期
721-724
,共4页
司马晋%张保%王保军%艾青%马鑫%张旭
司馬晉%張保%王保軍%艾青%馬鑫%張旭
사마진%장보%왕보군%애청%마흠%장욱
前列腺肿瘤%Notch1基因%Hes1基因%细胞迁移%细胞增殖
前列腺腫瘤%Notch1基因%Hes1基因%細胞遷移%細胞增殖
전렬선종류%Notch1기인%Hes1기인%세포천이%세포증식
Prostate neoplasms%Notch1 gene%Hes1 gene%Cell migration%Cell proliferation
目的 研究体外下调Notch1基因对前列腺癌细胞迁移和增殖能力的影响.方法 选取人前列腺癌PC-3细胞,分别转染Notch1的小干扰RNA(siRNA)序列(实验组)和阴性对照序列(阴性对照组),使用荧光定量PCR和蛋白质印迹技术分别检测转染后各组细胞中Notch1和其下游靶基因Hes1的表达.以Transwell小室法和MTS法分别检测细胞的迁移和增殖能力.结果 较之阴性对照组和空白对照组,实验组Notch1基因相对表达量显著降低(实验组3.960±0.510,阴性对照组36.097±1.941,空白对照组38.762±1.897),差异具有统计学意义(P＜ 0.01).在Notch1表达下调的同时,实验组中Hes1的表达也明显下降(实验组1.690±0.994,阴性对照组8.776±0.916,空白对照组9.803±1.001)(P＜0.01).迁移实验中,实验组细胞数为(657.867±27.610)个,明显多于阴性对照组的(158.533±18.263)个和空白对照组的(146.933±15.733)个,差异有统计学意义(P＜ 0.01).细胞增殖能力方面,开始检测时细胞增殖能力差异无统计学意义(P＞0.05),实验组细胞在24、48、72 h的吸光度(A)值均高于两对照组细胞(均P＜0.01).结论 下调Notch1基因可以促进前列腺癌PC-3细胞的迁移和增殖,这一效应可能是通过Notch1调节其靶基因Hes1实现的.
目的 研究體外下調Notch1基因對前列腺癌細胞遷移和增殖能力的影響.方法 選取人前列腺癌PC-3細胞,分彆轉染Notch1的小榦擾RNA(siRNA)序列(實驗組)和陰性對照序列(陰性對照組),使用熒光定量PCR和蛋白質印跡技術分彆檢測轉染後各組細胞中Notch1和其下遊靶基因Hes1的錶達.以Transwell小室法和MTS法分彆檢測細胞的遷移和增殖能力.結果 較之陰性對照組和空白對照組,實驗組Notch1基因相對錶達量顯著降低(實驗組3.960±0.510,陰性對照組36.097±1.941,空白對照組38.762±1.897),差異具有統計學意義(P＜ 0.01).在Notch1錶達下調的同時,實驗組中Hes1的錶達也明顯下降(實驗組1.690±0.994,陰性對照組8.776±0.916,空白對照組9.803±1.001)(P＜0.01).遷移實驗中,實驗組細胞數為(657.867±27.610)箇,明顯多于陰性對照組的(158.533±18.263)箇和空白對照組的(146.933±15.733)箇,差異有統計學意義(P＜ 0.01).細胞增殖能力方麵,開始檢測時細胞增殖能力差異無統計學意義(P＞0.05),實驗組細胞在24、48、72 h的吸光度(A)值均高于兩對照組細胞(均P＜0.01).結論 下調Notch1基因可以促進前列腺癌PC-3細胞的遷移和增殖,這一效應可能是通過Notch1調節其靶基因Hes1實現的.
목적 연구체외하조Notch1기인대전렬선암세포천이화증식능력적영향.방법 선취인전렬선암PC-3세포,분별전염Notch1적소간우RNA(siRNA)서렬(실험조)화음성대조서렬(음성대조조),사용형광정량PCR화단백질인적기술분별검측전염후각조세포중Notch1화기하유파기인Hes1적표체.이Transwell소실법화MTS법분별검측세포적천이화증식능력.결과 교지음성대조조화공백대조조,실험조Notch1기인상대표체량현저강저(실험조3.960±0.510,음성대조조36.097±1.941,공백대조조38.762±1.897),차이구유통계학의의(P＜ 0.01).재Notch1표체하조적동시,실험조중Hes1적표체야명현하강(실험조1.690±0.994,음성대조조8.776±0.916,공백대조조9.803±1.001)(P＜0.01).천이실험중,실험조세포수위(657.867±27.610)개,명현다우음성대조조적(158.533±18.263)개화공백대조조적(146.933±15.733)개,차이유통계학의의(P＜ 0.01).세포증식능력방면,개시검측시세포증식능력차이무통계학의의(P＞0.05),실험조세포재24、48、72 h적흡광도(A)치균고우량대조조세포(균P＜0.01).결론 하조Notch1기인가이촉진전렬선암PC-3세포적천이화증식,저일효응가능시통과Notch1조절기파기인Hes1실현적.
Objective To investigate the effect of Notch1 gene dowuregulated on migration and proliferation of human prostate cancer cell line PC-3.Methods The small interfering RNA (siRNA) targeted Notch1 gene or negative control sequences was transfected into PC-3 cells.The expression of Notch1 or Hes1 gene was detected by Real-Time PCR and Western Blot.Then ability of migration or proliferation was measured by Transwell assay or MTS Assay.Results Compared with negative control group (36.097±1.941) and untransfected group (38.762±1.897),Notch1 expression level in siRNA group (3.960±0.510) was significantly reduced (P ＜ 0.01).Meanwhile,Hes1 level in siRNA group was decreased,expression in three groups as follows:siRNA group was 1.690±0.994,negative control group was 8.776±0.916,untransfected group was 9.803±1.001 (P ＜ 0.01).In Transwell assay,the number of migration cells in siRNA group was 657.867±27.610,more than that in the negative control group (158.533±18.263) and untransfected group (146.933±15.733) (P ＜ 0.01).In MTS assay,there was no significant difference among three groups at 0 h point,however,siRNA group was significantly raised at the time points of 24,48 and 72 h (P ＜ 0.01).Conclusions Downregulation of Notch1 gene by transfection of the siRNA-Notch1 sequences significantly promoted ability of migration or proliferation in PC-3 cells,and the effect may be due to the down-regulation of Hes1 expression.
