RNA干扰沉默NF-κB p65基因表达抑制胰腺癌PANC1细胞增殖与侵袭的研究

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.

Silencing of NF-κB p65 expression by RNA interference inhibits proliferation and invasion of pancreatic carcinoma cell strain PANC1

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.