目的 探讨胰腺癌组织中微RNA-100的表达及其与胰腺癌患者临床病理因素的关系,以及微RNA-100在胰腺癌发生发展过程中的作用及其可能机制.方法 回顾性分析2013年1月至2014年3月贵州医科大学附属医院收治的17例胰腺癌患者的临床病理资料.收集患者手术切除标本进行分析.采用实时荧光定量PCR检测胰腺癌组织中微RNA-100的表达,分析其与胰腺癌患者临床病理因素的关系.采用慢病毒过表达载体(lv-微RNA-100)感染人胰腺癌MIA PaCa-2和CFPAC-1细胞,将MIA PaCa-2细胞分为M组和N组:M组转染lv-微RNA-100,N组转染对照慢病毒空载体(lv-control);将CFPAC-1细胞分为C组和D组:C组转染lv-微RNA-100,D组转染lv-control.利用实时荧光定量PCR检测MIA PaCa-2和CFPAC-1细胞中微RNA-100表达情况,利用流式细胞仪检测MIA PaCa-2和CFPAC-1细胞周期,利用Western blot检测MIA PaCa-2和CFPAC-1细胞中Cyclin D1蛋白表达情况.正态分布的计量资料以(x)±s表示,组间比较采用t检验.结果 实时荧光定量PCR检测:微RNA-100在胰腺癌组织中的相对表达量为0.046±0.020,低于癌旁组织的0.097±0.017,两者比较,差异有统计学意义(t=2.789,P＜0.05).不同肿瘤分化程度和肿瘤分期的胰腺癌患者,胰腺癌组织中微RNA-109表达水平比较,差异有统计学意义(t=2.563,2.135,P＜0.05).实时荧光定量PCR检测:微RNA-100在M组细胞中的相对表达量为0.097±0.012,高于N组的0.018±0.006,两组比较,差异有统计学意义(t=4.410,P＜0.05).微RNA-100在C组细胞中的相对表达量为0.084±0.021,高于D组的0.023±0.010,两组比较,差异有统计学意义(t=5.351,P＜0.05).流式细胞仪检测:M组细胞G0～G1期细胞百分比为45.3％±0.7％,N组细胞为30.6％±0.7％,两组比较,差异有统计学意义(t=5.564,P＜0.05).C组细胞G0～ G1期细胞百分比为58.8％±1.0％,D组细胞为42.6％±0.7％,两组比较,差异有统计学意义(t=7.771,P＜0.05).Westem blot检测:M组细胞Cyclin D1蛋白相对表达量为0.352±0.081,N组细胞为0.872±0.134,两组比较,差异有统计学意义(t=7.651,P＜0.05).C组细胞Cyclin D1蛋白相对表达量为0.410±0.121,D组细胞为0.979±0.232,两组比较,差异有统计学意义(t=8.712,P＜0.05).结论 微RNA-100在胰腺癌组织中表达下调,可能与肿瘤分化程度和肿瘤分期有关,其可能作用机制为通过抑制Cyclin D1蛋白的表达抑制胰腺癌细胞增殖.
目的 探討胰腺癌組織中微RNA-100的錶達及其與胰腺癌患者臨床病理因素的關繫,以及微RNA-100在胰腺癌髮生髮展過程中的作用及其可能機製.方法 迴顧性分析2013年1月至2014年3月貴州醫科大學附屬醫院收治的17例胰腺癌患者的臨床病理資料.收集患者手術切除標本進行分析.採用實時熒光定量PCR檢測胰腺癌組織中微RNA-100的錶達,分析其與胰腺癌患者臨床病理因素的關繫.採用慢病毒過錶達載體(lv-微RNA-100)感染人胰腺癌MIA PaCa-2和CFPAC-1細胞,將MIA PaCa-2細胞分為M組和N組:M組轉染lv-微RNA-100,N組轉染對照慢病毒空載體(lv-control);將CFPAC-1細胞分為C組和D組:C組轉染lv-微RNA-100,D組轉染lv-control.利用實時熒光定量PCR檢測MIA PaCa-2和CFPAC-1細胞中微RNA-100錶達情況,利用流式細胞儀檢測MIA PaCa-2和CFPAC-1細胞週期,利用Western blot檢測MIA PaCa-2和CFPAC-1細胞中Cyclin D1蛋白錶達情況.正態分佈的計量資料以(x)±s錶示,組間比較採用t檢驗.結果 實時熒光定量PCR檢測:微RNA-100在胰腺癌組織中的相對錶達量為0.046±0.020,低于癌徬組織的0.097±0.017,兩者比較,差異有統計學意義(t=2.789,P＜0.05).不同腫瘤分化程度和腫瘤分期的胰腺癌患者,胰腺癌組織中微RNA-109錶達水平比較,差異有統計學意義(t=2.563,2.135,P＜0.05).實時熒光定量PCR檢測:微RNA-100在M組細胞中的相對錶達量為0.097±0.012,高于N組的0.018±0.006,兩組比較,差異有統計學意義(t=4.410,P＜0.05).微RNA-100在C組細胞中的相對錶達量為0.084±0.021,高于D組的0.023±0.010,兩組比較,差異有統計學意義(t=5.351,P＜0.05).流式細胞儀檢測:M組細胞G0～G1期細胞百分比為45.3％±0.7％,N組細胞為30.6％±0.7％,兩組比較,差異有統計學意義(t=5.564,P＜0.05).C組細胞G0～ G1期細胞百分比為58.8％±1.0％,D組細胞為42.6％±0.7％,兩組比較,差異有統計學意義(t=7.771,P＜0.05).Westem blot檢測:M組細胞Cyclin D1蛋白相對錶達量為0.352±0.081,N組細胞為0.872±0.134,兩組比較,差異有統計學意義(t=7.651,P＜0.05).C組細胞Cyclin D1蛋白相對錶達量為0.410±0.121,D組細胞為0.979±0.232,兩組比較,差異有統計學意義(t=8.712,P＜0.05).結論 微RNA-100在胰腺癌組織中錶達下調,可能與腫瘤分化程度和腫瘤分期有關,其可能作用機製為通過抑製Cyclin D1蛋白的錶達抑製胰腺癌細胞增殖.
목적 탐토이선암조직중미RNA-100적표체급기여이선암환자림상병리인소적관계,이급미RNA-100재이선암발생발전과정중적작용급기가능궤제.방법 회고성분석2013년1월지2014년3월귀주의과대학부속의원수치적17례이선암환자적림상병리자료.수집환자수술절제표본진행분석.채용실시형광정량PCR검측이선암조직중미RNA-100적표체,분석기여이선암환자림상병리인소적관계.채용만병독과표체재체(lv-미RNA-100)감염인이선암MIA PaCa-2화CFPAC-1세포,장MIA PaCa-2세포분위M조화N조:M조전염lv-미RNA-100,N조전염대조만병독공재체(lv-control);장CFPAC-1세포분위C조화D조:C조전염lv-미RNA-100,D조전염lv-control.이용실시형광정량PCR검측MIA PaCa-2화CFPAC-1세포중미RNA-100표체정황,이용류식세포의검측MIA PaCa-2화CFPAC-1세포주기,이용Western blot검측MIA PaCa-2화CFPAC-1세포중Cyclin D1단백표체정황.정태분포적계량자료이(x)±s표시,조간비교채용t검험.결과 실시형광정량PCR검측:미RNA-100재이선암조직중적상대표체량위0.046±0.020,저우암방조직적0.097±0.017,량자비교,차이유통계학의의(t=2.789,P＜0.05).불동종류분화정도화종류분기적이선암환자,이선암조직중미RNA-109표체수평비교,차이유통계학의의(t=2.563,2.135,P＜0.05).실시형광정량PCR검측:미RNA-100재M조세포중적상대표체량위0.097±0.012,고우N조적0.018±0.006,량조비교,차이유통계학의의(t=4.410,P＜0.05).미RNA-100재C조세포중적상대표체량위0.084±0.021,고우D조적0.023±0.010,량조비교,차이유통계학의의(t=5.351,P＜0.05).류식세포의검측:M조세포G0～G1기세포백분비위45.3％±0.7％,N조세포위30.6％±0.7％,량조비교,차이유통계학의의(t=5.564,P＜0.05).C조세포G0～ G1기세포백분비위58.8％±1.0％,D조세포위42.6％±0.7％,량조비교,차이유통계학의의(t=7.771,P＜0.05).Westem blot검측:M조세포Cyclin D1단백상대표체량위0.352±0.081,N조세포위0.872±0.134,량조비교,차이유통계학의의(t=7.651,P＜0.05).C조세포Cyclin D1단백상대표체량위0.410±0.121,D조세포위0.979±0.232,량조비교,차이유통계학의의(t=8.712,P＜0.05).결론 미RNA-100재이선암조직중표체하조,가능여종류분화정도화종류분기유관,기가능작용궤제위통과억제Cyclin D1단백적표체억제이선암세포증식.
Objective To investigate the effects and mechanism of expression of microRNA-100 in the tissues of pancreatic cancer and its relationship with clinicopathological factor.Methods The clinical data of 17 patients with pancreatic cancer who were admitted to the Affiliated Hospital of Guizhou Medical University between January 2013 and March 2014 were retrospectively analyzed.The surgical specimens were collected for study.The expression of microRNA-100 in the pancreatic tissues were detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and its relationship with clinicopathological factors was analyzed.The MIA PaCa-2 cells and CFPAC-1 cells were infected by lv-microRNA-100.MIA PaCa-2 cells were divided into the M group (microRNA-100 were infected) and N group (empty vectors were infected),and CFPAC-1 cells were divided into the C group (microRNA-100 were infected) and D group (empty vectors were infected).The expressions of microRNA-100 in the CFPAC-1 cells and MIA PaCa-2 cells were detected by RT-PCR and cell cycles were detected by flow cytometry.The expressions of Cyclin D1 protein in the CFPAC-1 cells and MIA PaCa-2 cells were detected by Western blot.Measurement data with normal distribution were presented as (x) ± s and comparison among groups were done by t test.Results The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the pancreatic cancer tissues was 0.046 ± 0.020,which was significantly different from 0.097 ± 0.017 in the adjacent tissues (t =2.789,P ＜ 0.05).There were significant differences between the tumor differentiation degree and tumor staging in patients with pancreatic cancer (t =2.563,2.135,P ＜0.05).The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the M group was 0.097 ±0.012,which was significantly higher than 0.018 ± 0.006 in the N group (t =4.410,P ＜ 0.05).The relative quantitative expression of microRNA-100 in the C group was 0.084 ± 0.021,which was significantly higher than 0.023 ± 0.010 in the D group (t =5.351,P ＜ 0.05).The results of flow cytometry showed that the percentage of cells between G0-G1 were 45.3％ ±0.7％ in the M group and 30.6％ ±0.7％ in the N group,with a significant difference (t =5.564,P ＜ 0.05).The percentage of cells between G0-G1 were 58.8％ ± 1.0％ in the C group and 42.6％ ± 0.7％ in the D group,with a significant difference (t =7.771,P ＜ 0.05).The results of Western blot showed that the relative quantitative expression of Cyclin D1 protein were 0.352 ± 0.081 in the M group and 0.872 ± 0.134 in the N group,with a significant difference (t =7.651,P ＜ 0.05).The relative quantitative expression of Cyclin D1 protein was 0.410 ± 0.121 in the C group,which was significantly different from 0.979 ± 0.232 in the D group (t =8.712,P ＜ 0.05).Conclusion Low expression of microRNA-100 in pancreatic cancer tissues may be correlated with tumor differentiation degree and tumor staging,it can inhibit tumor cells proliferation by reducing expression of Cyclin D1 protein.
