β_1整合素反义寡核苷酸联合顺铂治疗人卵巢癌裸鼠移植瘤的研究

Experimental study of β_1 integrin antisense oligodeonucleotide in combination with cisplatin for human ovarian carcinoma xenograft in nude mice

目的：探讨以β1整合素反义寡核苷酸（ASODN）治疗人卵巢癌裸鼠皮下移植瘤的可行性.方法：常规体外培养SKOV3细胞,采用皮下注射法建立移植瘤裸鼠动物模型.32只荷瘤鼠随机分为ASODN组（A组）、ASODN联合顺铂（DDP）组（A＋D组）、DDP组和0.9％氯化钠注射液对照组（NS组）,每组8只,分组给药.以脂质体包裹的β1整合素ASODN直接移植瘤内注射,观察肿瘤生长情况,测瘤体积并计算抑瘤率.采用反转录-聚合酶链反应（RT-PCR）和免疫组织化学方法分别检测β1整合素mRNA的表达.结果：A组肿瘤体积和抑瘤率分别为（316.10 ±21.77） mm3和48.15％,与NS组比较抑瘤率较高,肿瘤生长缓慢（P＜0.01）.而A＋D组肿瘤体积和抑瘤率分别为（178.70±40.67） mm3和70.37％,与DDP组、A组及NS组差异均有统计学意义（P ＜0.05 ～P＜0.01）.RT-PCR和免疫组织化学检测,A组和A＋D组肿瘤组织中β1整合素mRNA的表达均明显下调（P＜0.01）.结论：单用β1整合素ASDON或联用DDP均可有效抑制人卵巢癌裸鼠皮下移植瘤组织的生长,可能与其特异性下调β1整合素基因表达有关.特异性靶向β1整合素ASODN可用于卵巢癌的辅助治疗.

Objective：To explore the feasibility of antisense oligodeoxynucleotide （ASODN） targeting β1 integrin gene for treatment of human ovarian cancer in nude mice. Methods：Routinely cultured SKOV3 cells were subcutaneously implanted in the nude mice to establish xenograft animal model. The 32 tumor-bearing mice were randomly divided into ASODN group （ A group）, ASODN combined with DDP group （ A ＋ D group）, DDP group and physiological saline control group （ NS group） （ n = 8 in each ）. Drug treatment was started after randomization. After 131integrin ASODN mediated by cytofectin was directly injected into the xenograft, the volume and weight of the tumor mass were detected, and then the tumor growth inhibitory rate was calculated. Reverse transcription-polymerase chain reaction（RT-PCR） and immunochemohistology assay were used to detect the expression level of [31integrin mRNA. Results： In the A group, the tumor volume and tumor anticancer efficacy were （316.10 ± 21.77 ）mm3 and 48.15 %, respectively. The tumor growth inhibitory rate and tumor volume were significantly different between the NS group and the A group（P 〈0. 01 ）. In the A ＋ D group,the tumor volume reduced to （178.70 ± 40.67）mm^3 and the tumor anticancer efficacy was enhanced to 70.37 % ,which were significantly different compared with those of the DDP group and other controls （ P 〈 0.05 to P 〈 0.01 ）. Significant down-regulation of β1 integrin mRNA expression in the tumor tissues of the A group and A ＋ D group was detected by RT-PCR and immunochemohistology assay（P 〈 0.01 ）. Conclusions： Singly use of β1 integrin ASODN or combined with DDP can effectively inhibit the growth of human ovarian carcinoma xenograft in nude mice by direct intra-tumoral injection. The anticancer efficacy may be associated with the down regulation of β1 integrin expression. ASODN targeting β1 integrin gene can be a supportive therapy to the ovarian cancer.