中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2015年
4期
233-236
,共4页
王楠%杨剑峰%贾振宇%何杨%许春芳
王楠%楊劍峰%賈振宇%何楊%許春芳
왕남%양검봉%가진우%하양%허춘방
胰腺肿瘤%肿瘤侵袭%丝氨酸蛋白酶类%基质金属蛋白酶9
胰腺腫瘤%腫瘤侵襲%絲氨痠蛋白酶類%基質金屬蛋白酶9
이선종류%종류침습%사안산단백매류%기질금속단백매9
Pancreatic neoplasms%Neoplasm invasiveness%Serine proteases%Matrix metalloproteinase 9
目的 探讨matriptase基因沉默对胰腺癌细胞株SW1990侵袭力的影响.方法 采用脂质体法将不同浓度的靶向matriptase的siRNA (Ma-siRNA)转染人胰腺癌SW1990细胞株,以转染阴性对照的siRNA (NC-siRNA)作为对照组.采用荧光定量PCR法和蛋白质印迹法检测各组细胞matriptasemRNA和蛋白的表达水平,采用Transwell小室检测细胞侵袭能力,明胶酶谱法检测细胞matriptase及MMP-9的活性.结果 NC-siRNA组及12.5、25、50 nmol/L Ma-siRNA组SW1990细胞matriptase mRNA相对表达量分别为1.000、0.417±0.006、0.233±0.068、0.221±0.092,蛋白表达量分别为0.736±0.066、0.498±0.036、0.341 ±0.118、0.239±0.050,Ma-siRNA各组细胞matriptase mRNA及蛋白表达量均显著低于NC-siRNA组,差异具有统计学意义(P值均＜0.01);matriptase活性分别为1.501 ±0.165、1.211±0.265、0.645 ±0.165、0.620±0.003,MMP-9活性分别为0.929±0.260、0.484±0.364、0.352±0.113、0.346±0.121,其中25、50 nmol/L Ma-siRNA组细胞的matriptase及MMP-9活性较NC-siRNA组显著下降,差异有统计学意义(P值＜0.05或＜0.01).NC-siRNA组的穿膜细胞数为(256±1)个/200倍视野,25 nmo]/L Ma-siRNA组为(109±3)个/200倍视野,25 nmol/L Ma-siRNA组细胞侵袭力较对照组下降(57.4±5.4)％.结论 抑制matriptase基因表达可降低胰腺癌SW1990细胞的侵袭能力,其机制可能与matriptase及MMP-9酶活性下降有关.
目的 探討matriptase基因沉默對胰腺癌細胞株SW1990侵襲力的影響.方法 採用脂質體法將不同濃度的靶嚮matriptase的siRNA (Ma-siRNA)轉染人胰腺癌SW1990細胞株,以轉染陰性對照的siRNA (NC-siRNA)作為對照組.採用熒光定量PCR法和蛋白質印跡法檢測各組細胞matriptasemRNA和蛋白的錶達水平,採用Transwell小室檢測細胞侵襲能力,明膠酶譜法檢測細胞matriptase及MMP-9的活性.結果 NC-siRNA組及12.5、25、50 nmol/L Ma-siRNA組SW1990細胞matriptase mRNA相對錶達量分彆為1.000、0.417±0.006、0.233±0.068、0.221±0.092,蛋白錶達量分彆為0.736±0.066、0.498±0.036、0.341 ±0.118、0.239±0.050,Ma-siRNA各組細胞matriptase mRNA及蛋白錶達量均顯著低于NC-siRNA組,差異具有統計學意義(P值均＜0.01);matriptase活性分彆為1.501 ±0.165、1.211±0.265、0.645 ±0.165、0.620±0.003,MMP-9活性分彆為0.929±0.260、0.484±0.364、0.352±0.113、0.346±0.121,其中25、50 nmol/L Ma-siRNA組細胞的matriptase及MMP-9活性較NC-siRNA組顯著下降,差異有統計學意義(P值＜0.05或＜0.01).NC-siRNA組的穿膜細胞數為(256±1)箇/200倍視野,25 nmo]/L Ma-siRNA組為(109±3)箇/200倍視野,25 nmol/L Ma-siRNA組細胞侵襲力較對照組下降(57.4±5.4)％.結論 抑製matriptase基因錶達可降低胰腺癌SW1990細胞的侵襲能力,其機製可能與matriptase及MMP-9酶活性下降有關.
목적 탐토matriptase기인침묵대이선암세포주SW1990침습력적영향.방법 채용지질체법장불동농도적파향matriptase적siRNA (Ma-siRNA)전염인이선암SW1990세포주,이전염음성대조적siRNA (NC-siRNA)작위대조조.채용형광정량PCR법화단백질인적법검측각조세포matriptasemRNA화단백적표체수평,채용Transwell소실검측세포침습능력,명효매보법검측세포matriptase급MMP-9적활성.결과 NC-siRNA조급12.5、25、50 nmol/L Ma-siRNA조SW1990세포matriptase mRNA상대표체량분별위1.000、0.417±0.006、0.233±0.068、0.221±0.092,단백표체량분별위0.736±0.066、0.498±0.036、0.341 ±0.118、0.239±0.050,Ma-siRNA각조세포matriptase mRNA급단백표체량균현저저우NC-siRNA조,차이구유통계학의의(P치균＜0.01);matriptase활성분별위1.501 ±0.165、1.211±0.265、0.645 ±0.165、0.620±0.003,MMP-9활성분별위0.929±0.260、0.484±0.364、0.352±0.113、0.346±0.121,기중25、50 nmol/L Ma-siRNA조세포적matriptase급MMP-9활성교NC-siRNA조현저하강,차이유통계학의의(P치＜0.05혹＜0.01).NC-siRNA조적천막세포수위(256±1)개/200배시야,25 nmo]/L Ma-siRNA조위(109±3)개/200배시야,25 nmol/L Ma-siRNA조세포침습력교대조조하강(57.4±5.4)％.결론 억제matriptase기인표체가강저이선암SW1990세포적침습능력,기궤제가능여matriptase급MMP-9매활성하강유관.
Objective To investigate the effect of the down regulation of matriptase expression on invasion of human pancreatic cancers cells SW1990.Methods Small interfering RNA targeting at matriptase (Ma-siRNA) was transfected into human pancreatic cancers SW1990 cells,and nonsense siRNA (NC-siRNA) group was used as control.Real time PCR assay and Western blot were used to detect the expression of matriptase mRNA and protein.Transwell assay was used to examine the invasion ability of cancer cells.The enzymatic activity of matriptase and MMP-9 was determined by gelatin zymography assay.Results The expression level of matriptase mRNA in NC-siRNA group,12.5,25,50 nmol/L Ma-siRNA group were 1.000,0.417 ± 0.006,0.233 ± 0.068,0.221 ± 0.092;and the protein expression of matriptase were 0.736 ± 0.066,0.498 ± 0.036,0.341 ± 0.118,0.239 ± 0.050,respectively.The matriptase mRNA and protein expression in Ma-siRNA groups was significantly lower than those in NC-siRNA group,and the difference between the two groups was statistically significant (P ＜ 0.05).The enzymatic activity of matriptase were 1.501 ±0.165,1.211 ±0.265,0.645 ±0.165,0.620 ±0.003,and the enzymatic activity of MMP-9 were 0.929 ± 0.260,0.484 ± 0.364,0.352 ± 0.113,0.346 ± 0.121,and the enzymatic activity of matriptase and MMP-9 in 25,50 nmol/L Ma-siRNA groups was significantly lower than that in NC-siRNA group,and the difference was statistically significant (P ＜ 0.05 or ＜ 0.01).The number of transmembrane cell was (256 ± 1)/per 200 power field,and it was (109 ± 3)/per 200 power field in 25 nmol/L Ma-siRNA group,and the invasion ability of the cells in 25 nmol/L Ma-siRNA group was decreased by (57.4 ± 5.4) ％ when compared with that of control group.Conclusions Down-regulation of matriptase inhibits invasion ability of pancreatic cancer SW1990 cells,and this result may be due to the down regulated enzymatic activity of matriptase and MMP-9.
