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
84-87
,共4页
王殿忠%谢敏%潘一明%朱海涛
王殿忠%謝敏%潘一明%硃海濤
왕전충%사민%반일명%주해도
胰腺肿瘤%RNA干扰%NF-κB%ICAM-1
胰腺腫瘤%RNA榦擾%NF-κB%ICAM-1
이선종류%RNA간우%NF-κB%ICAM-1
Pancreatic neoplasms%RNA interference%NF-κB%ICAM-1
目的 运用RNA干扰技术阻断人胰腺癌细胞株PANC1的NF-κB p65表达,观察该基因沉默后对细胞增殖能力及体外侵袭能力的影响.方法 采用阳离子脂质体LipofectamineTM 2000将化学合成的人NF-κB p65的小干扰RNA(siRNA)转染人胰腺癌细胞PANC1作为siRNA组,另设无同源性siRNA转染细胞的阴性对照组和蒸馏水空白对照组.实验重复6次.应用RT-PCR检测转染细胞NF-κB p65 mRNA与细胞间黏附分子-1(ICAM-1)mRNA的表达.MTT结合克隆形成实验测定细胞生长抑制率.Matrigel细胞侵袭实验测定细胞体外侵袭能力.结果 siRNA组、阴性对照组和空白对照组NF-κB p65 mRNA相对表达量分别为0.227±0.045、0.381±0.038和0.404±0.031;ICAM-1 mRNA相对表达量分别为0.597±0.083、0.983±0.068和1.027±0.098;细胞生长抑制率(72 h)分别为18.3%、2.3%和0;克隆形成率分别为(14.1±3.1)%、(24.5±2.1)%和(27.2±2.6)%;侵袭细胞数分别为80.25±6.35、123.83±8.80和127.68±9.23.siRNA组均明显低于2个对照组(P＜0.01).结论 NF-κB p65的RNA干扰能抑制胰腺癌PANC1细胞NF-κB p65 mRNA和ICAM-1 mRNA的表达,抑制细胞的生长和侵袭.
目的 運用RNA榦擾技術阻斷人胰腺癌細胞株PANC1的NF-κB p65錶達,觀察該基因沉默後對細胞增殖能力及體外侵襲能力的影響.方法 採用暘離子脂質體LipofectamineTM 2000將化學閤成的人NF-κB p65的小榦擾RNA(siRNA)轉染人胰腺癌細胞PANC1作為siRNA組,另設無同源性siRNA轉染細胞的陰性對照組和蒸餾水空白對照組.實驗重複6次.應用RT-PCR檢測轉染細胞NF-κB p65 mRNA與細胞間黏附分子-1(ICAM-1)mRNA的錶達.MTT結閤剋隆形成實驗測定細胞生長抑製率.Matrigel細胞侵襲實驗測定細胞體外侵襲能力.結果 siRNA組、陰性對照組和空白對照組NF-κB p65 mRNA相對錶達量分彆為0.227±0.045、0.381±0.038和0.404±0.031;ICAM-1 mRNA相對錶達量分彆為0.597±0.083、0.983±0.068和1.027±0.098;細胞生長抑製率(72 h)分彆為18.3%、2.3%和0;剋隆形成率分彆為(14.1±3.1)%、(24.5±2.1)%和(27.2±2.6)%;侵襲細胞數分彆為80.25±6.35、123.83±8.80和127.68±9.23.siRNA組均明顯低于2箇對照組(P＜0.01).結論 NF-κB p65的RNA榦擾能抑製胰腺癌PANC1細胞NF-κB p65 mRNA和ICAM-1 mRNA的錶達,抑製細胞的生長和侵襲.
목적 운용RNA간우기술조단인이선암세포주PANC1적NF-κB p65표체,관찰해기인침묵후대세포증식능력급체외침습능력적영향.방법 채용양리자지질체LipofectamineTM 2000장화학합성적인NF-κB p65적소간우RNA(siRNA)전염인이선암세포PANC1작위siRNA조,령설무동원성siRNA전염세포적음성대조조화증류수공백대조조.실험중복6차.응용RT-PCR검측전염세포NF-κB p65 mRNA여세포간점부분자-1(ICAM-1)mRNA적표체.MTT결합극륭형성실험측정세포생장억제솔.Matrigel세포침습실험측정세포체외침습능력.결과 siRNA조、음성대조조화공백대조조NF-κB p65 mRNA상대표체량분별위0.227±0.045、0.381±0.038화0.404±0.031;ICAM-1 mRNA상대표체량분별위0.597±0.083、0.983±0.068화1.027±0.098;세포생장억제솔(72 h)분별위18.3%、2.3%화0;극륭형성솔분별위(14.1±3.1)%、(24.5±2.1)%화(27.2±2.6)%;침습세포수분별위80.25±6.35、123.83±8.80화127.68±9.23.siRNA조균명현저우2개대조조(P＜0.01).결론 NF-κB p65적RNA간우능억제이선암PANC1세포NF-κB p65 mRNA화ICAM-1 mRNA적표체,억제세포적생장화침습.
Objective To investigate the inhibitory effect of proliferation and invasiveness on PANC1 cell by knockdown of NF-κB P65 expression through RNA interference. Methods Chemically synthesized small interference RNA (siRNA) directed against human NF-κB p65 was transfected into pancreatic carcinoma cell (PANC1) by using cationic liposome LipofectamineTM 2000 as the transfection agent. A non-silencing siRNA was used as the negative control and distilled water was used as the blank control. The tests repeated for six times. The expression of NF-KB p65 and ICAM-1 gone was detected by RT-PCR. The effect of cell proliferation and invasive ability was evaluated by MTF, clony assay, Matrigel invasive tests. Results The relative expression levels of NF-κB p65 mRNA in siRNA, the negative control and the blank control group were 0.227 ±0.045, O. 381 ± 0. 038 and 0. 404 ± 0. 031, respectively; the relative expression levels of ICAM-1mRNA were 0.597 ± 0. 083, 0.983 ± 0.068 and 1. 027 ± 0.098, respectively; the inhibitory rate of the cells at 72h were 18.3%, 2.3% and 0, respectively; the colony forming efficiency were ( 14.1 ± 3.1 ) %, ( 24.5±2.1)% and (27.2 ±2.6)%, respectively; the number of invasive cells were 80.25 ±6.35, 123.83 ±8.80 and 127.68 ± 9.23, respectively; the difference between the siRNA group and two control groups was significant (P ＜ 0. 01 ). Conclusions Down-regulatiag the expression of NF-κB p65 gene by RNAi may inhibit proliferation and invasiveness in pancreatic carcinoma cell strain PANC1.
