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目的:探讨Ad-ING4人膀胱癌裸鼠移植瘤的生长抑制作用及其分子机制。方法:1Ad空载体腺病毒(Ad-GFP)、Ad-ING4单基因腺病毒(Ad-ING4)分别经QBI-293A细胞感染多轮扩增后获得高效价重组病毒;2将浓度为4×107个/ml的T24膀胱癌细胞悬液在裸鼠右前肢腋下皮下注射,建立T24人膀胱癌裸鼠移植瘤模型;3将15只荷瘤裸鼠随机分为阴性对照组(PBS组)、Ad-GFP组、Ad-ING4组,3组均使用瘤体内注射干预用药,隔日1次,共6次。观测瘤体生长情况,测量肿瘤长径和短径,用公式:肿瘤体积=ab2/2(a为长径,b为短径),计算肿瘤体积,治疗3周后将裸鼠脱颈处死,摘瘤称重,用10%甲醛溶液固定标本,石蜡包埋,进行HE染色和免疫组织化学检测,并按Weidner的方法检测微血管密度(MVD)。结果:治疗后Ad-ING4组肿瘤重量为(0.93±0.06)g,分别与PBS组(1.73±0.05)g及Ad-GFP组(1.69±0.06)g相比,差异有统计学意义(P<0.05)。AdING4的抑瘤率达45%。Ad-ING4组肿瘤组织中细胞凋亡相关因子Bax和caspase-3的表达明显上调,Bcl-2的表达明显下调,肿瘤血管形成相关因子CD34表达下调,MVD降低(P<0.05)。结论:1Ad-ING4可以显著抑制膀胱癌移植瘤生长。2Ad-ING4的抑瘤机制可能和激活细胞凋亡及抑制血管形成有关。
Objective: To investigate the growth inhibitory effect and its molecular mechanism of Ad-ING4 human bladder carcinoma xenografts in nude mice. Methods: Ad-GFP and Ad-ING4 adenovirus Ad-ING4 were transfected into QBI-293A cells for multiple rounds of amplification to obtain high titer recombinant virus. / Ml T24 bladder cancer cell suspension in the right forelimb of the nude mice subcutaneously injected subcutaneously to establish a T24 human bladder cancer xenograft model in nude mice; 3 15 nude mice were randomly divided into negative control group (PBS group) Ad-GFP group, Ad-ING4 group, 3 groups were used intratumoral injection intervention medication, every other day, a total of 6 times. Observe the growth of the tumor and measure the long and short diameter of the tumor. Calculate the tumor volume by the formula: tumor volume = ab2 / 2 (a is the long diameter and b is the short diameter). After 3 weeks of treatment, The tumors were weighed, fixed with 10% formaldehyde solution and embedded in paraffin for HE staining and immunohistochemistry. Microvascular density (MVD) was measured by Weidner’s method. Results: The tumor weight of Ad-ING4 group was (0.93 ± 0.06) g after treatment, which was significantly different from PBS group (1.73 ± 0.05) g and Ad-GFP group (1.69 ± 0.06) g respectively 0.05). The inhibition rate of AdING4 is 45%. The expression of Bax and caspase-3, the expression of Bcl-2 and the expression of CD34 in Ad-ING4 group were significantly decreased (P <0.05). Conclusion: 1Ad-ING4 can significantly inhibit the growth of transplanted bladder cancer. The antitumor mechanism of 2Ad-ING4 may be related to the activation of apoptosis and the inhibition of angiogenesis.