中华胰腺病学杂志
中華胰腺病學雜誌
중화이선병학잡지
CHINESE JOURNAL OF PANCREATOLOGY
2008年
2期
81-83
,共3页
弭延斌%郭晓钟%刘峰%徐建华%田宏%夏春莲%吴开春%樊代明
弭延斌%郭曉鐘%劉峰%徐建華%田宏%夏春蓮%吳開春%樊代明
미연빈%곽효종%류봉%서건화%전굉%하춘련%오개춘%번대명
胰腺肿瘤%RNA,小分子干扰%KAI1基因%细胞系
胰腺腫瘤%RNA,小分子榦擾%KAI1基因%細胞繫
이선종류%RNA,소분자간우%KAI1기인%세포계
Pancreatic neoplasms%RNA,small interfering%KAI1%Cell line
目的 应用小分子干扰RNA(small interference RNA,siRNA)干扰KAI1基因在人胰腺癌T3细胞系的表达,为基因治疗胰腺癌打下基础.方法 针对KAI1基因CD82片段序列设计A、B、C、D 4个siRNA靶序列,构建针对KAI1基因(CD82)含siRNA片段的慢病毒载体.以脂质体2000转染T3细胞,测定病毒滴度.以空载体、含不同靶序列siRNAd1载体的病毒感染T3细胞(MOI=5),采用实时PCR方法检测CD82mRNA的表达.结果 正常对照组、空载体对照组、A靶序列组、B靶序列组、C靶序列组和D靶序列组CD82 mRNA表达量分别为1.398 ±0.242、1.311 ±0.048、0.664±0.093、0.345 ±0.032、0.641 ±0.049和0.147±0.049,各靶序列组的表达量明显低于空载体对照组(P＜0.01).结论 应用RNAi可有效抑制T3细胞KAI1基因CD82片段的表达.
目的 應用小分子榦擾RNA(small interference RNA,siRNA)榦擾KAI1基因在人胰腺癌T3細胞繫的錶達,為基因治療胰腺癌打下基礎.方法 針對KAI1基因CD82片段序列設計A、B、C、D 4箇siRNA靶序列,構建針對KAI1基因(CD82)含siRNA片段的慢病毒載體.以脂質體2000轉染T3細胞,測定病毒滴度.以空載體、含不同靶序列siRNAd1載體的病毒感染T3細胞(MOI=5),採用實時PCR方法檢測CD82mRNA的錶達.結果 正常對照組、空載體對照組、A靶序列組、B靶序列組、C靶序列組和D靶序列組CD82 mRNA錶達量分彆為1.398 ±0.242、1.311 ±0.048、0.664±0.093、0.345 ±0.032、0.641 ±0.049和0.147±0.049,各靶序列組的錶達量明顯低于空載體對照組(P＜0.01).結論 應用RNAi可有效抑製T3細胞KAI1基因CD82片段的錶達.
목적 응용소분자간우RNA(small interference RNA,siRNA)간우KAI1기인재인이선암T3세포계적표체,위기인치료이선암타하기출.방법 침대KAI1기인CD82편단서렬설계A、B、C、D 4개siRNA파서렬,구건침대KAI1기인(CD82)함siRNA편단적만병독재체.이지질체2000전염T3세포,측정병독적도.이공재체、함불동파서렬siRNAd1재체적병독감염T3세포(MOI=5),채용실시PCR방법검측CD82mRNA적표체.결과 정상대조조、공재체대조조、A파서렬조、B파서렬조、C파서렬조화D파서렬조CD82 mRNA표체량분별위1.398 ±0.242、1.311 ±0.048、0.664±0.093、0.345 ±0.032、0.641 ±0.049화0.147±0.049,각파서렬조적표체량명현저우공재체대조조(P＜0.01).결론 응용RNAi가유효억제T3세포KAI1기인CD82편단적표체.
Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.
