论文部分内容阅读
目的:研究重组p53腺病毒(recombinant adenovirus-p53,rAd-p53)在体内、外对肺腺癌H1299细胞(野生型p53基因缺失)生长的抑制作用,观察rAd-p53尾静脉注射治疗肺腺癌的可行性。方法:MTT法检测rAd-p53对H1299细胞增殖的抑制作用。rAd-p53以感染复数(multiplicity of infection,MOI)=500感染H1299细胞,24 h后RT-PCR检测H1299细胞中p53mRNA表达,72 h后Western blotting检测P53蛋白的表达、流式细胞术检测H1299细胞凋亡。H1299细胞皮下接种BALB/c裸鼠,建立裸鼠肺腺癌模型,尾静脉注射rAd-p53,观察肿瘤生长情况,绘制肿瘤生长曲线。结果:rAd-p53以MOI=500感染H1299细胞,24 h后有野生型p53 mRNA转录,72 h后有P53蛋白表达;且rAd-p53感染可明显抑制H1299细胞增殖,72 h时,rAd-p53组细胞增殖比显著低于对照组(2.8±0.4 vs 6.1±0.5,P<0.05)。感染rAd-p53后,随时间增加H1299细胞凋亡率呈上升趋势,48 h时rAd-p53组细胞凋亡率显著高于对照组,[(27.6±0.05)%vs(4.9±0.09)%,P<0.01]。成功建立H1299细胞荷瘤裸鼠模型,尾静脉注射rAd-p53 2周后移植瘤体积显著小于对照组[(0.875±0.253)vs(0.479±0.215)cm3,P<0.05]。结论:rAd-p53感染可上调H1299细胞P53蛋白的表达,抑制H1299细胞增殖、促进其凋亡,并且尾静脉注射rAd-p53可明显抑制H1299细胞裸鼠移植瘤的生长。
Objective: To study the inhibitory effect of recombinant adenovirus-p53 (rAd-p53) on the growth of lung adenocarcinoma H1299 cells (wild-type p53 gene) in vivo and in vitro and to observe the effect of rAd-p53 tail vein injection on lung adenocarcinoma Feasibility. Methods: The inhibitory effect of rAd-p53 on proliferation of H1299 cells was detected by MTT assay. H1299 cells were infected with rAd-p53 at a multiplicity of infection (MOI) = 500. The expression of p53 mRNA in H1299 cells was detected by RT-PCR at 24 h and the expression of P53 protein at 72 h after transfection. Flow cytometry was used to detect H1299 cells Apoptosis. BALB / c nude mice were inoculated subcutaneously in H1299 cells to establish a model of lung adenocarcinoma in nude mice. The rAd-p53 was injected into the caudal vein to observe the tumor growth and to draw the tumor growth curve. Results: rAd-p53 infected H1299 cells with MOI = 500. After 24 h, wild-type p53 mRNA was transcribed and P53 protein was expressed after 72 h. RAd-p53 infection significantly inhibited the proliferation of H1299 cells. After 72 h, rAd- Group cell proliferation was significantly lower than the control group (2.8 ± 0.4 vs 6.1 ± 0.5, P <0.05). After being infected with rAd-p53, the apoptosis rate of H1299 cells increased with time, and the apoptosis rate of rAd-p53 group was significantly higher than that of control group at 48 h [(27.6 ± 0.05)% vs (4.9 ± 0.09)%, P <0.01]. Successfully established H1299 tumor-bearing nude mice model, tail vein injection of rAd-p53 2 weeks after the tumor volume was significantly smaller than the control group [(0.875 ± 0.253) vs (0.479 ± 0.215) cm3, P <0.05]. Conclusion: The rAd-p53 infection can up-regulate the expression of P53 protein in H1299 cells, inhibit the proliferation and promote the apoptosis of H1299 cells, and rAd-p53 can significantly inhibit the growth of H1299 cells transplanted in nude mice.