中华泌尿外科杂志
中華泌尿外科雜誌
중화비뇨외과잡지
CHINESE JOURNAL OF UROLOGY
2014年
3期
226-229
,共4页
司马晋%张保%马鑫%王保军%艾青%张旭
司馬晉%張保%馬鑫%王保軍%艾青%張旭
사마진%장보%마흠%왕보군%애청%장욱
膀胱癌%Numb基因%基质金属蛋白酶-9%侵袭%迁移
膀胱癌%Numb基因%基質金屬蛋白酶-9%侵襲%遷移
방광암%Numb기인%기질금속단백매-9%침습%천이
Bladder neoplasms%Numb Gene%Matrix metalloproteinase-9%Invasion%Migration
目的 研究Numb基因对膀胱癌细胞侵袭和迁移能力的影响及相关机制.方法 2013年6月我们使用Numb-ORF表达质粒和pCMV6-Entry空载体质粒转染人膀胱癌细胞T24,作为实验组和阴性对照组,无质粒转染的人膀胱癌细胞T24作为空白对照组,采用荧光定量PCR和蛋白质印迹技术检测各组的Numb和基质金属蛋白酶-9(MMP-9)的表达情况,采用细胞划痕实验和Transwell小室法分别检测细胞的迁移和侵袭能力.结果 荧光定量PCR检测显示,与阴性对照组(△CT值5.871±0.685)和空白对照组(△CT值5.618±0.597)比较,实验组中Numb表达(△CT值1.655±0.432)显著升高(P＜0.01).实验组的MMP-9表达(△CT值8.423±0.202)低于阴性对照组(△CT值7.294±0.138)和空白对照组(△CT值7.220±0.118),差异有统计学意义(P＜0.01).划痕实验中,12h的划痕修复比实验组为0.525±0.037,阴性对照组为0.693±0.034,空白对照组为0.701±0.038,组间比较差异有统计学意义(P=0.004).侵袭实验中,实验组的穿膜细胞数为(12.6±3.2)个,明显少于阴性对照组[(130.8±9.2)个]和空白对照组[(132.2±10.6)个],差异有统计学意义(P＜0.05).结论 上调Numb基因可以降低人膀胱癌细胞T24的侵袭和迁移能力,其机制可能是通过Numb负性调节MMP-9实现的.
目的 研究Numb基因對膀胱癌細胞侵襲和遷移能力的影響及相關機製.方法 2013年6月我們使用Numb-ORF錶達質粒和pCMV6-Entry空載體質粒轉染人膀胱癌細胞T24,作為實驗組和陰性對照組,無質粒轉染的人膀胱癌細胞T24作為空白對照組,採用熒光定量PCR和蛋白質印跡技術檢測各組的Numb和基質金屬蛋白酶-9(MMP-9)的錶達情況,採用細胞劃痕實驗和Transwell小室法分彆檢測細胞的遷移和侵襲能力.結果 熒光定量PCR檢測顯示,與陰性對照組(△CT值5.871±0.685)和空白對照組(△CT值5.618±0.597)比較,實驗組中Numb錶達(△CT值1.655±0.432)顯著升高(P＜0.01).實驗組的MMP-9錶達(△CT值8.423±0.202)低于陰性對照組(△CT值7.294±0.138)和空白對照組(△CT值7.220±0.118),差異有統計學意義(P＜0.01).劃痕實驗中,12h的劃痕脩複比實驗組為0.525±0.037,陰性對照組為0.693±0.034,空白對照組為0.701±0.038,組間比較差異有統計學意義(P=0.004).侵襲實驗中,實驗組的穿膜細胞數為(12.6±3.2)箇,明顯少于陰性對照組[(130.8±9.2)箇]和空白對照組[(132.2±10.6)箇],差異有統計學意義(P＜0.05).結論 上調Numb基因可以降低人膀胱癌細胞T24的侵襲和遷移能力,其機製可能是通過Numb負性調節MMP-9實現的.
목적 연구Numb기인대방광암세포침습화천이능력적영향급상관궤제.방법 2013년6월아문사용Numb-ORF표체질립화pCMV6-Entry공재체질립전염인방광암세포T24,작위실험조화음성대조조,무질립전염적인방광암세포T24작위공백대조조,채용형광정량PCR화단백질인적기술검측각조적Numb화기질금속단백매-9(MMP-9)적표체정황,채용세포화흔실험화Transwell소실법분별검측세포적천이화침습능력.결과 형광정량PCR검측현시,여음성대조조(△CT치5.871±0.685)화공백대조조(△CT치5.618±0.597)비교,실험조중Numb표체(△CT치1.655±0.432)현저승고(P＜0.01).실험조적MMP-9표체(△CT치8.423±0.202)저우음성대조조(△CT치7.294±0.138)화공백대조조(△CT치7.220±0.118),차이유통계학의의(P＜0.01).화흔실험중,12h적화흔수복비실험조위0.525±0.037,음성대조조위0.693±0.034,공백대조조위0.701±0.038,조간비교차이유통계학의의(P=0.004).침습실험중,실험조적천막세포수위(12.6±3.2)개,명현소우음성대조조[(130.8±9.2)개]화공백대조조[(132.2±10.6)개],차이유통계학의의(P＜0.05).결론 상조Numb기인가이강저인방광암세포T24적침습화천이능력,기궤제가능시통과Numb부성조절MMP-9실현적.
Objective To study the effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24,and its related mechanism.Methods In June 2013,the plasmids including Numb-ORF plasmid and its blank vector pCMV6-Entry were transfected into T24 cells,and set the blank control as usual.The expressions of Numb and MMP-9 (Matrix metalloproteinase-9) were detected by Real-Time PCR and Western Blot.Then T24 cell invasion and migration were measured respectively by Transwell assay and Scratch Wound Healing Assay.Results Compared to negative control group,which transfected with blank-vector plasmid and untransfected group,Numb expression level in Numb-ORF group was significantly higher (P＜0.01).Meanwhile,MMP-9 level in Numb-ORF group was decreased,△ CT-value as follows:Numb-ORF group was 8.423±0.202,negative control group was 7.294±0.138,untransfected group was 7.220±0.118 (P＜0.01).In Scratch Wound Healing Assay,the repair ratio of Numb-ORF group was less than negative control group and untransfected group.The repair ratio in 12 hours as follow:Numb-ORF group was 0.525±0.037,negative control group was 0.693±0.034,untransfected group was 0.701 ±0.038(P=0.004).In Transwell assay,the number of invasion cells in Numb-ORF group was 12.6±3.2,less than in control group (130.8±9.2) and untransfected group (132.2± 10.6) (P＜0.05).Conclusions Overexpression of Numb by transfection significantly decreases the capability of T24 cell invasion and migration.This suppressed effect may be due to the down-regulation of MMP-9 expression in human bladder cancer.
