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目的探讨Survivin反义寡核苷酸联合血管内皮生长因子(VEGF)反义寡核苷酸在裸鼠体内对人肺腺癌细胞A549的生长抑制作用。方法将脂质体包裹的ASODN混悬液瘤旁注射移植有A549肿瘤细胞的裸鼠,并相应地把裸鼠分为5组:空白对照组、脂质体对照组、VEGF处理组、Survivin处理组,Survivin-VEGF联合处理组。观察各组肿瘤的生长情况,并分别以Western blot、real-time PCR检测肿瘤组织Survivin及VEGF各自的表达情况,用免疫组化法检测Cyclin B1的蛋白表达和微血管密度计数(CD34染色)。结果 Survivin-VEGF ASODN联合处理组较各组的生长情况明显减慢(P<0.05);Western blot和real-time PCR结果显示Survivin-VEGF联合处理组中Survivin和VEGF蛋白及RNA的表达较各组均受到明显抑制(P<0.05);Survivin-VEGF联合处理组显著降低了肿瘤组织微血管密度计数(MVD)(10.27±1.75),与各单反义寡核苷酸处理组[Survivin处理组(13.27±2.19),VEGF处理组(11.40±1.18)]之间的差异具有统计学意义(P<0.05);Survivin-VEGF联合处理组停滞于G2/M期的细胞(23.17±5.45)也较Survivin处理组(12.54±6.67)和VEGF处理组(18.39±6.87)明显增多(P<0.05)。结论联合使用VEGF反义寡核苷酸和Survivin寡核苷酸对肿瘤的抑制作用优于单用VEGF反义寡核苷酸或Survivin反义寡核苷酸。
Objective To investigate the inhibitory effect of Survivin antisense oligonucleotide combined with vascular endothelial growth factor (VEGF) antisense oligonucleotide on human lung adenocarcinoma A549 cells in nude mice. Methods Nude mice bearing A549 tumor cells were transplanted with liposome-coated ASODN suspension. Nude mice were divided into 5 groups: blank control group, liposome control group, VEGF treatment group and Survivin treatment group Group, Survivin-VEGF combined treatment group. The growth of each group was observed. The expression of Survivin and VEGF were detected by Western blot and real-time PCR respectively. The protein expression of Cyclin B1 and the count of microvessel density (CD34) were detected by immunohistochemistry. Results Compared with the control group, the growth of Survivin-VEGF ASODN group was significantly slower than that of the control group (P <0.05). The results of Western blot and real-time PCR showed that the expressions of Survivin and VEGF protein and RNA in Survivin- (P <0.05). Survivin-VEGF combined treatment significantly reduced the microvessel density (MVD) of tumor tissue (10.27 ± 1.75), compared with the single-antisense oligonucleotide treatment group [Survivin treatment group 2.19), VEGF group (11.40 ± 1.18)] (P <0.05). Compared with Survivin group, the cells arrested in G2 / M phase in Survivin-VEGF group (23.17 ± 5.45) (12.54 ± 6.67) and VEGF treatment group (18.39 ± 6.87) (P <0.05). Conclusion The combination of VEGF antisense oligonucleotide and Survivin oligonucleotide has better antitumor activity than VEGF antisense oligonucleotide or Survivin antisense oligonucleotide.