中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2012年
9期
602-606
,共5页
黄李雅%陈平%张树贤%刘磊%盛露露%袁耀宗
黃李雅%陳平%張樹賢%劉磊%盛露露%袁耀宗
황리아%진평%장수현%류뢰%성로로%원요종
趋化因子CX3CL1%转染%胰腺肿瘤%细胞系,肿瘤%RNA,小分子干扰
趨化因子CX3CL1%轉染%胰腺腫瘤%細胞繫,腫瘤%RNA,小分子榦擾
추화인자CX3CL1%전염%이선종류%세포계,종류%RNA,소분자간우
Chemokine CX3CL1%Transfection%Pancreatic neoplasms%Cell line,tumor%RNA,small interfering
目的 探讨阻断趋化因子Fractalkine(FKN)对胰腺癌细胞株SW-1990和PNAC-1生物学功能的影响.方法 腺病毒作为载体包装FKN-小干扰RNA(siRNA),转染人胰腺癌细胞株SW-1990和PNAC-1.应用克隆形成实验、MTT法和细胞侵袭实验检测FKN-siRNA转染与否、胰腺癌细胞增殖和侵袭力的变化.Western印迹法和RT-PCR检测转染FKN-siRNA的胰腺癌细胞FKN、TNF-α和IL-6蛋白和mRNA的表达.统计学处理采用单因素方差分析.结果 FKN-siRNA转染人胰腺癌细胞株SW-1990和PNAC-1后,与对照组的克隆数[分别为(1.97％±0.25％)和(1.77％±0.25％)]和阴性FKN-siRNA组的克隆数[分别为(2.10％±0.30％)和(1.97％±0.25％)]相比,细胞克隆增大,克隆数[分别为(5.27％±0.35％)和(4.60％±0.30％)]增多,差异均有统计学意义(F=113.51、103.86,P值均＜0.05).MTT实验48 h和72 h后,转入FKN-siRNA的转染人胰腺癌细胞株SW-1990和PNAC-1吸光度(48 h时分别为1.28±0.07和1.19±0.14;72 h时分别为1.49±0.11和1.52±0.16)高于对照组([48 h时分别为0.80±0.03和0.74±0.11;72 h时分别为0.89±0.03和0.93±0.04)和阴性FKN-siRNA组(48 h时分别为0.85±0.02和0.76±0.05;72 h时分别为0.89±0.02和1.07±0.09),差异均有统计学意义(F=83.80、71.99、17.19、23.51,P值均＜0.05).细胞侵袭实验中,转入FKN-siRNA的细胞侵袭力强于对照组和阴性FKN-siRNA组,差异有统计学意义(F=37.37、9.08,P值均＜0.05).FKN-siRNA转染后,SW-1990和PNAC-1细胞中,FKN蛋白(F=118.93、88.62)和mRNA(F=47.91、72.59)表达水平下降,同时TNF-α和IL-6的蛋白(FTNF-α=112.90、77.88,FIL-6=165.27、286.49)和mRNA(FTNFα=47.93、45.19,FIL-6=36.41、23.67)表达水平增高,差异均有统计学意义(P均＜0.05).结论 应用siRNA技术静默趋化因子FKN的功能后,胰腺癌细胞株的增殖活性和侵袭力增强,表明FKN可能抑制了胰腺癌细胞的生物学活性.
目的 探討阻斷趨化因子Fractalkine(FKN)對胰腺癌細胞株SW-1990和PNAC-1生物學功能的影響.方法 腺病毒作為載體包裝FKN-小榦擾RNA(siRNA),轉染人胰腺癌細胞株SW-1990和PNAC-1.應用剋隆形成實驗、MTT法和細胞侵襲實驗檢測FKN-siRNA轉染與否、胰腺癌細胞增殖和侵襲力的變化.Western印跡法和RT-PCR檢測轉染FKN-siRNA的胰腺癌細胞FKN、TNF-α和IL-6蛋白和mRNA的錶達.統計學處理採用單因素方差分析.結果 FKN-siRNA轉染人胰腺癌細胞株SW-1990和PNAC-1後,與對照組的剋隆數[分彆為(1.97％±0.25％)和(1.77％±0.25％)]和陰性FKN-siRNA組的剋隆數[分彆為(2.10％±0.30％)和(1.97％±0.25％)]相比,細胞剋隆增大,剋隆數[分彆為(5.27％±0.35％)和(4.60％±0.30％)]增多,差異均有統計學意義(F=113.51、103.86,P值均＜0.05).MTT實驗48 h和72 h後,轉入FKN-siRNA的轉染人胰腺癌細胞株SW-1990和PNAC-1吸光度(48 h時分彆為1.28±0.07和1.19±0.14;72 h時分彆為1.49±0.11和1.52±0.16)高于對照組([48 h時分彆為0.80±0.03和0.74±0.11;72 h時分彆為0.89±0.03和0.93±0.04)和陰性FKN-siRNA組(48 h時分彆為0.85±0.02和0.76±0.05;72 h時分彆為0.89±0.02和1.07±0.09),差異均有統計學意義(F=83.80、71.99、17.19、23.51,P值均＜0.05).細胞侵襲實驗中,轉入FKN-siRNA的細胞侵襲力彊于對照組和陰性FKN-siRNA組,差異有統計學意義(F=37.37、9.08,P值均＜0.05).FKN-siRNA轉染後,SW-1990和PNAC-1細胞中,FKN蛋白(F=118.93、88.62)和mRNA(F=47.91、72.59)錶達水平下降,同時TNF-α和IL-6的蛋白(FTNF-α=112.90、77.88,FIL-6=165.27、286.49)和mRNA(FTNFα=47.93、45.19,FIL-6=36.41、23.67)錶達水平增高,差異均有統計學意義(P均＜0.05).結論 應用siRNA技術靜默趨化因子FKN的功能後,胰腺癌細胞株的增殖活性和侵襲力增彊,錶明FKN可能抑製瞭胰腺癌細胞的生物學活性.
목적 탐토조단추화인자Fractalkine(FKN)대이선암세포주SW-1990화PNAC-1생물학공능적영향.방법 선병독작위재체포장FKN-소간우RNA(siRNA),전염인이선암세포주SW-1990화PNAC-1.응용극륭형성실험、MTT법화세포침습실험검측FKN-siRNA전염여부、이선암세포증식화침습력적변화.Western인적법화RT-PCR검측전염FKN-siRNA적이선암세포FKN、TNF-α화IL-6단백화mRNA적표체.통계학처리채용단인소방차분석.결과 FKN-siRNA전염인이선암세포주SW-1990화PNAC-1후,여대조조적극륭수[분별위(1.97％±0.25％)화(1.77％±0.25％)]화음성FKN-siRNA조적극륭수[분별위(2.10％±0.30％)화(1.97％±0.25％)]상비,세포극륭증대,극륭수[분별위(5.27％±0.35％)화(4.60％±0.30％)]증다,차이균유통계학의의(F=113.51、103.86,P치균＜0.05).MTT실험48 h화72 h후,전입FKN-siRNA적전염인이선암세포주SW-1990화PNAC-1흡광도(48 h시분별위1.28±0.07화1.19±0.14;72 h시분별위1.49±0.11화1.52±0.16)고우대조조([48 h시분별위0.80±0.03화0.74±0.11;72 h시분별위0.89±0.03화0.93±0.04)화음성FKN-siRNA조(48 h시분별위0.85±0.02화0.76±0.05;72 h시분별위0.89±0.02화1.07±0.09),차이균유통계학의의(F=83.80、71.99、17.19、23.51,P치균＜0.05).세포침습실험중,전입FKN-siRNA적세포침습력강우대조조화음성FKN-siRNA조,차이유통계학의의(F=37.37、9.08,P치균＜0.05).FKN-siRNA전염후,SW-1990화PNAC-1세포중,FKN단백(F=118.93、88.62)화mRNA(F=47.91、72.59)표체수평하강,동시TNF-α화IL-6적단백(FTNF-α=112.90、77.88,FIL-6=165.27、286.49)화mRNA(FTNFα=47.93、45.19,FIL-6=36.41、23.67)표체수평증고,차이균유통계학의의(P균＜0.05).결론 응용siRNA기술정묵추화인자FKN적공능후,이선암세포주적증식활성화침습력증강,표명FKN가능억제료이선암세포적생물학활성.
Objective To explore the effects of Fractalkine (FKN) on the biological functions of human pancreatic cancer cell lines SW-1990 and PNAC-1.Methods Adenovirus mediated FKN-small interfering RNA (siRNA) was transfected into human pancreatic cancer cell lines SW 1990 and PNAC-1.The differences in proliferation and invasion ability between before and after FKN-siRNA transfection were determined by clone formation assay,MTT assay and cells invasion assay.After FKN-siRNA transfection,the expression of FKN,tumor necrosis factor (TNF)-α and interleukin (IL)-6 at protein and mRNA level in human pancreatic cancer cell were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR).The data were analyzed by one way analysis of variance.Results After human pancreatic cancer cell lines SW-1990 and PNAC-1 transfected with FKN-siRNA,the clone numbers (5.27 ％ ± 0.35 ％ and 4.60 ％ ± 0.30％ ) increased compared with those of control group ( 1.97％ ±0.25％ and 1.77％ ± 0.25％ ) and negative FKN-siRNA group (2.10％±0.30％ and 1.97％±0.25％),and the difference was statistically significant (F=113.51,103.86; both P＜0.05).The clone size was also enlarged.After human pancreatic cancer cell lines SW-1990 and PNAC-1 transfected with FKN-siRNA for 48 hours and 72 hours,the MTT test results showed the absorbance value (48 h:1.28±0.07 and 1.19±0.14; 72 h:1.49±0.11 and 1.52±0.16) was higher than that of control group (48 h:0.80±0.03 and 0.74±0.11;72 h:0.89±0.03 and 0.93±0.04) and negative FKN-siRNA group (48 h:0.85±0.02 and 0.76±0.05; 72 h:0.89±0.02 and 1.07±0.09),and the difference was statistically significant (F=83.80,71.99,17.19,23.51; all P＜0.05).The invasion ability assay showed that the invasion ability of FKN-siRNA transfected cells was stronger than that of control group and negative FKN-siRNA group,and the difference was statistically significant (F=37.37,9.08; both P＜0.05).After FKN-siRNA transfection,the expression of FKN at protein and mRNA level in SW-1990 and PNAC-1 cell line decreased (protein:F=118.93 and 88.62,mRNA:F=47.91 and 72.59),at the same time the expression of TNF-α and IL-6 at protein and mRNA level increased (protein:FTNF-α =112.90 and 77.88,FIL-6 =165.27 and 286.49,mRNA:FTNF-α ==47.93 and 45.19,FIL-6 =36.41 and 23.67),and the differences were statistically significant (all P values＜0.05).Conclusion With siRNA technology to silent FKN function,the proliferation and invasion ability of pancreatic cancer cell lines increased,which indicated FKN might inhibit certain biological functions of pancreatic cancer cells.
