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目的构建人pVITRO2-RhoB重组质粒,探讨pVITRO2-RhoB重组质粒在体内外的抗肿瘤效果及其联合低剂量顺铂对小鼠A549肺癌的治疗作用。方法构建pVITRO2-RhoB重组质粒,将其转染A549细胞后,观察它在细胞中的表达情况以及对A549细胞的抑制作用。建立小鼠A549肺癌模型,用1,2-二油酰-3-三甲胺基丙烷(DOTAP)+胆固醇(CHOL)脂质体包裹pVITRO2-RhoB重组质粒制备脂质体质粒DNA复合体即Lip-RhoB。将30只荷瘤鼠分别分为空白组、脂质体-pVITRO2复合物(Lip-null)组、脂质体-RhoB复合物(Lip-RhoB)组、低剂量顺铂(DDP)组和联合治疗(RhoB+DDP)组。RhoB尾静脉给药,低剂量顺铂腹腔给药,观察各组肿瘤体积变化及小鼠的生存期,原位瘤进行TUNEL染色,测定凋亡率。结果成功构建了pVITRO2-RhoB重组质粒,该重组质粒在体外抑制了A549细胞生长并促进了该细胞的凋亡;pVITRO2-RhoB重组质粒治疗裸鼠A549肺癌皮下肿瘤模型后,荷瘤鼠肿瘤体积增长减慢,肿瘤凋亡细胞增加(P<0.05);基因联合低剂量顺铂治疗后,与Lip-RhoB组和DDP组相比能显著抑制荷瘤鼠肿瘤生长(P<0.05),延长荷瘤鼠生存期(P<0.05),提高肿瘤细胞原位凋亡(P<0.05)。结论 RhoB基因具有明显的体内外抗肿瘤效果及协同化疗药物的抗肿瘤作用,因此RhoB基因联合低剂量顺铂可能是一种有效的肺癌治疗新方法。
Objective To construct the recombinant plasmid pVITRO2-RhoB and investigate the antitumor effect of pVITRO2-RhoB recombinant plasmid in vitro and in vivo and the therapeutic effect of pVITRO2-RhoB combined with low-dose cisplatin on A549 lung cancer in mice. Methods The recombinant plasmid pVITRO2-RhoB was constructed and transfected into A549 cells. The expression of pVITRO2-RhoB in A549 cells was observed and its inhibitory effect on A549 cells was also observed. A mouse model of A549 lung cancer was established, and the plasmid pVITRO2-RhoB was constructed by encapsulating pVITRO2-RhoB recombinant plasmid with 1,2-dioleoyl-3-trimethylaminopropane (DOTAP) + cholesterol (CHOL) RhoB. Thirty tumor-bearing mice were divided into blank group, Lip-null group, Lip-RhoB group, low-dose cisplatin (DDP) Treatment (RhoB + DDP) group. RhoB tail vein administration, low-dose cisplatin intraperitoneal administration, changes in tumor volume and survival time were observed in each group, TUNEL staining in situ tumor, the rate of apoptosis was measured. Results The recombinant plasmid pVITRO2-RhoB was constructed successfully. The recombinant plasmid could inhibit the growth of A549 cells and promote the apoptosis of A549 cells in vitro. After the recombinant plasmid pVITRO2-RhoB was used to treat the A549 lung cancer subcutaneous tumor model in nude mice, the tumor volume increased (P <0.05). Compared with Lip-RhoB group and DDP group, the gene therapy combined with low-dose cisplatin could significantly inhibit tumor growth of tumor-bearing mice (P <0.05) and prolong tumorigenicity (P <0.05), and increased the apoptosis of tumor cells in situ (P <0.05). Conclusion RhoB gene has obvious anti-tumor effect in vitro and in vivo and anti-tumor effect of chemotherapeutic drugs. Therefore, RhoB gene combined with low-dose cisplatin may be a new effective treatment for lung cancer.