siRNA靶向沉默HIF-1a和RBM5过表达对前列腺癌PC3细胞生长抑制作用的体外研究

In vitro study of SiRNA targeted silencing of hif-1aand RBM5overexpression on growth inhibition of prostate cancer PC3 cells

目的观察采用RNA干扰抑制前列腺癌(PCa)PC3细胞中缺氧诱导因子-1a(HIF-1a)和上调RBM5表达水平对PC3细胞增殖和凋亡的影响。方法运用PEI/RBM5/HA-HP载体(RBM5组)及siHIF-1a载体(si HIF-1a组)分别或联合(RBM5+siHIF-1a组)转染PC3细胞,以未转染细胞(空白对照组)和空载体细胞(阴性对照组)作为比较。采用噻唑蓝比色(MTT)法检测各组细胞增殖情况;流式细胞仪(FCM)检测各组细胞周期变化;原位末端标记(TUNEL)法检测各组细胞凋亡指数。结果RBM5组、si HIF-1a组和RBM5+si HIF-1a的联合组均能不同程度的抑制PC3细胞增殖,使细胞周期停滞和促进细胞凋亡,但联合组作用更大,效果更好。联合组细胞增殖率是50.9%,与RBM5组的77.3%和siHIF-1a组的74.8%比较,差异有统计学意义(P<0.05),与空白对照组的100%和阴性对照组的98.6%比较,差异有显著统计学意义(P<0.01)。联合组对细胞周期和细胞凋亡的影响也明显优于单纯用药的两组,组间差异显著(P<0.05),与空白对照组和阴性对照组比较,差异更显著(P<0.01)。结论si RNA靶向沉默癌基因HIF-1a或PEI/RBM5/HA-HP载体增强抑癌基因RBM5表达这两种方法均可抑制前列腺癌PC3细胞生长,但两者联合应用效果更好。

Objective To observe the effect of RNA interference on the proliferation and apoptosis of PCa PC3cells by inhibiting the expression of hypoxia inducion-1a(HIF-1a)and up-regulating the expression of RBM5.Methods PC3cells were transfected with PEI/RBM5/HA-HP vector(RBM5group)and siHIF-1avector(si HIF-1agroup),respectively or in combination(RBM5+siHIF-1agroup).Untransfected cells(blank control group)and unloaded cells(negative control group)were compared.Cell cycle changes were detected by flow cytometry(FCM).TUNEL assay was used to detect the apoptosis index.Results RBM5group,siHIF-1agroup and RBM5+siHIF-1acombined group can inhibit PC3cell proliferation to varying degrees,make cell cycle arrest and promote apoptosis,but the combined group has a greater effect and better effect.The cell proliferation rate of the combined group was 50.9%,while that of the RBM5group was 77.3%and that of the siHIF-1agroup was 74.8%,with statistically significant difference(P<0.05),and the difference was statistically significant(P<0.01)compared with that of the blank control group(100%)and the negative control group(98.6%).The effect of the combined group on cell cycle and apoptosis was also significantly better than that of the two groups treated alone,and the difference between the two groups(P<0.05)was more significant than that of the blank control group and the negative control group(P<0.01).Conclusion siRNA-targeted silencing of oncogene HIF-1aor PEI/RBM5/HA-HP vector to enhance the expression of tumor suppressor gene RBM5can inhibit the growth of prostate cancer PC3cells,but the combined application of the two methods is better.