中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2014年
6期
366-369
,共4页
冯慧%王亚雷%陈熹%张辰宇%袁耀宗%姚玮艳
馮慧%王亞雷%陳熹%張辰宇%袁耀宗%姚瑋豔
풍혜%왕아뢰%진희%장신우%원요종%요위염
胰腺肿瘤%微RNAs%微囊泡
胰腺腫瘤%微RNAs%微囊泡
이선종류%미RNAs%미낭포
Pancreatic neoplasms%MicroRNAs%Microvesicle
目的 寻找胰腺癌循环微小核糖核酸(miRNA)的载体.方法 常规培养、传代胰腺癌细胞SW1990、BxPC3,采集6例胰腺癌患者及6例健康体检者(对照组)的血清.应用梯度离心方法获取血清和细胞培养上清中的微囊泡(MV).采用蛋白质印迹法检测RNA诱导沉默复合体的核心元件Ago2蛋白和MV的标志性蛋白CD63,采用荧光定量PCR方法检测MV包裹的和游离的miRNA.结果 胰腺癌细胞株培养上清的MV中含有Ago2、CD63蛋白.胰腺癌组、对照组的血清及MV中均存在miR-20a、miR-21、miR-24、miR-25、miR-191、miR-483-5p,但以MV包裹的量较多,且胰腺癌组MV包裹的miRNA量与对照组不完全一致,其中miR-20a、miR-24、miR-191分别为对照组的(2.93±0.29)、(2.73±0.46)、(2.39±0.51)倍,差异均有统计学意义(F值分别为75.97、25.80、12.94,P＜0.05或＜0.01).结论 MV是胰腺癌循环miRNA的主要载体.
目的 尋找胰腺癌循環微小覈糖覈痠(miRNA)的載體.方法 常規培養、傳代胰腺癌細胞SW1990、BxPC3,採集6例胰腺癌患者及6例健康體檢者(對照組)的血清.應用梯度離心方法穫取血清和細胞培養上清中的微囊泡(MV).採用蛋白質印跡法檢測RNA誘導沉默複閤體的覈心元件Ago2蛋白和MV的標誌性蛋白CD63,採用熒光定量PCR方法檢測MV包裹的和遊離的miRNA.結果 胰腺癌細胞株培養上清的MV中含有Ago2、CD63蛋白.胰腺癌組、對照組的血清及MV中均存在miR-20a、miR-21、miR-24、miR-25、miR-191、miR-483-5p,但以MV包裹的量較多,且胰腺癌組MV包裹的miRNA量與對照組不完全一緻,其中miR-20a、miR-24、miR-191分彆為對照組的(2.93±0.29)、(2.73±0.46)、(2.39±0.51)倍,差異均有統計學意義(F值分彆為75.97、25.80、12.94,P＜0.05或＜0.01).結論 MV是胰腺癌循環miRNA的主要載體.
목적 심조이선암순배미소핵당핵산(miRNA)적재체.방법 상규배양、전대이선암세포SW1990、BxPC3,채집6례이선암환자급6례건강체검자(대조조)적혈청.응용제도리심방법획취혈청화세포배양상청중적미낭포(MV).채용단백질인적법검측RNA유도침묵복합체적핵심원건Ago2단백화MV적표지성단백CD63,채용형광정량PCR방법검측MV포과적화유리적miRNA.결과 이선암세포주배양상청적MV중함유Ago2、CD63단백.이선암조、대조조적혈청급MV중균존재miR-20a、miR-21、miR-24、miR-25、miR-191、miR-483-5p,단이MV포과적량교다,차이선암조MV포과적miRNA량여대조조불완전일치,기중miR-20a、miR-24、miR-191분별위대조조적(2.93±0.29)、(2.73±0.46)、(2.39±0.51)배,차이균유통계학의의(F치분별위75.97、25.80、12.94,P＜0.05혹＜0.01).결론 MV시이선암순배miRNA적주요재체.
Objective To investigate the circulating microRNAs carrier in pancreatic cancer.Methods Pancreatic cancer cell lines SW1990 and BxPC3 were routinely cultured and serum of 6 patients with pancreatic cancer and 6 healthy subjects (control group) were collected.Serum and pancreatic cancer cell line supernatant microvesicles (MV) were obtained by gradient centrifugation.The expression of Ago2,CD63 was detected by Western blotting,the expression of microRNA in microvesicle section and microvesicle-free section in serum was detected by using quantitative PCR method.Results The supernatant MV of pancreatic cancer cell lines expressed Ago2,CD63 protein,and these MV carried different microRNAs in different cell lines.In the serum of pancreatic cancer and control group,miR-20a,miR-21,miR-24,miR-25,miR-191,miR-483-5p were detected,but the quantity was relatively higher in MV section,and the expression of microRNAs in pancreatic cancer's MV was inconsistent with that of control group.The expression of miR-20a,miR-24,miR-191 in pancreatic cancer group was (2.93 ± 0.29),(2.73 ± 0.46),(2.39 ± 0.51) times as much as those in control group,and the difference between the two groups was statistically significant (F =75.97,25.80,12.94,P ＜ 0.05 or ＜ 0.01).Conclusions The main circulating microRNAs carrier in pancreatic cancer is microvesicle.
