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目的观察腺病毒载体介导的抑癌基因LRIG1对前列腺癌细胞(PC-3细胞)增殖力、凋亡等生物学特性的影响。方法从前列腺增生组织获取目的基因LRIG1并将其与腺病毒骨架质粒pAd/PL-DEST包装,获取正确的重组腺病毒质粒pAD-LRIG1并扩增后,提取得到高滴度pAD-LRIG1(1×108PFU/mL)。将pAD-LRIG1按感染复数(MOI)=10加入PC-3细胞中,并设置空白对照,培养48h后,收集细胞并采用RT-PCR和Western Blot检测PC-3细胞中LRIG1的表达;将pAD-LRIG1按MOI=0、5、20(分别为对照组、低组、高组)分别加入PC-3细胞中,培养48h后,采用BrdU法检测细胞的增殖率,采用流式细胞仪检测细胞的凋亡率。结果通过RT-PCR和Western Blot检测,在PC-3细胞中加入pAD-LRIG1,能使LRIG1在核酸和蛋白水平表达;通过BrdU检测,对照组、低组、高组的细胞增殖率分别为(58.6±5.2)%、(41.3±4.7)%、(31.6±3.8)%,差异有统计学意义(P<0.05);通过流式细胞仪检测,对照组、低组、高组的细胞凋亡率分别为(3.8±0.6)%、(12.6±1.5)%、(18.2±3.1)%,差异有统计学意义(P<0.05)。结论腺病毒载体介导的抑癌基因LRIG1能增强PC-3细胞中LRIG1的表达,LRIG1基因表达上调后能有效抑制PC-3细胞的增殖率和增加细胞的凋亡率,为前列腺癌的进一步研究提供了理论依据。
Objective To observe the effect of adenovirus vector-mediated suppressor gene LRIG1 on the biological characteristics of prostate cancer cells (PC-3 cells) such as proliferation and apoptosis. Methods The target gene LRIG1 was obtained from prostatic hyperplasia tissues and packaged with the adenoviral backbone plasmid pAd / PL-DEST. The correct recombinant adenovirus plasmid pAD-LRIG1 was obtained and amplified. The high titer pAD-LRIG1 (1 × 108 PFU / mL). PC-3 cells were transfected with pAD-LRIG1 at multiplicity of infection (MOI) = 10, and blank control was set. After 48h incubation, cells were collected and the expression of LRIG1 in PC-3 cells was detected by RT-PCR and Western Blot. -LRIG1 was added to PC-3 cells at MOI = 0,5,20 (control group, low group and high group respectively). After cultured for 48h, the proliferation rate of cells was detected by BrdU method. The cells were detected by flow cytometry The rate of apoptosis. Results The expression of LRIG1 was detected by RT-PCR and Western Blot in PC-3 cells with pAD-LRIG1. The cell proliferation rates of control group, low group and high group were ( 58.6 ± 5.2)%, (41.3 ± 4.7)% and (31.6 ± 3.8)%, respectively. There was significant difference between the two groups (P <0.05). Flow cytometry The rates were (3.8 ± 0.6)%, (12.6 ± 1.5)% and (18.2 ± 3.1)%, respectively, with statistical significance (P <0.05). Conclusion Adenovirus vector-mediated suppressor gene LRIG1 can enhance the expression of LRIG1 in PC-3 cells. Upregulation of LRIG1 gene can effectively inhibit the proliferation rate of PC-3 cells and increase the apoptosis rate of PC-3 cells, Research provides a theoretical basis.