Department of Gynecology and Obstetrics, Key Laboratory of Obstetric and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital, State Key Laboratory of Biotherapy and Cancer Center, Sichuan University, Chengdu 610041, Sichuan, PR China.
Oncol Rep. 2012 Aug;28(2):668-76. doi: 10.3892/or.2012.1853. Epub 2012 Jun 6.
Currently, great interest is focused on the anti-neoplastic effects of CXC chemokine ligand 10 (IP-10/CXCL10). IP-10 has shown significant antitumor and anti-metastatic properties via immunological, antiangiogenic and anti-neoplastic mechanisms. However, very few studies on the antitumor activity of IP-10 in human ovarian cancer have been reported. The use of polymeric nanoparticles to deliver functional genes intraperitoneally holds much promise as an effective therapy for ovarian cancer. In our study, a recombinant plasmid expressing IP-10 (pVITRO-IP-10) was constructed, and biodegradable cationic heparin-polyethyleneimine (HPEI) nanogels were prepared to deliver pVITRO-IP-10 into SKOV3 human ovarian cancer cells. Transfection efficiency was detected by expression profiling of green fluorescent protein. The expression of IP-10 was determined using RT-PCR and western blot analysis. In vitro, cell proliferation was evaluated by MTT assay. Apoptosis was examined by Hoechst33258/PI staining and flow cytometry assays. The effect on the inhibition of angiogenesis was evaluated by tube formation assay using human umbilical vein endothelial cells (HUVECs). Moreover, a SKOV3 intraperitoneal ovarian carcinomatosis model was established to investigate the antitumor activity of HPEI+pVITRO-IP-10 complexes in nude mice. Tumor weights were evaluated during the treatment course. Cell proliferation and apoptosis were evaluated by Ki-67 immunochemical staining and TUNEL assay, and the antiangiogenic effect of pVITRO-IP-10 was assessed by CD31 immunochemical staining and alginate-encapsulated tumor cell assay. pVITRO-IP-10 was efficiently transfected into SKOV3 cells by HPEI nanogels. Intraperitoneal administration of HPEI+pVITRO-IP-10 complexes led to effective growth inhibition of ovarian cancer, in which tumor weight decreased by ~69.92% in the treatment group compared with that in the empty vector control group. Meanwhile, decreased cell proliferation, increased tumor cell apoptosis and reduction in angiogenesis were observed in the HPEI+pVITRO-IP-10 group compared with those in the control groups. These results indicated that HPEI nanogel delivery of pVITRO-IP-10 may be of value in the treatment against human ovarian cancer.
目前，人们对 CXC 趋化因子配体 10（IP-10/CXCL10）的抗肿瘤作用非常感兴趣。IP-10 通过免疫、抗血管生成和抗肿瘤机制表现出显著的抗肿瘤和抗转移特性。然而，关于 IP-10 在人卵巢癌中的抗肿瘤活性的研究非常少。使用聚合物纳米颗粒经腹腔内递送达功能性基因作为卵巢癌有效治疗的方法具有很大的前景。在我们的研究中，构建了表达 IP-10 的重组质粒（pVITRO-IP-10），并制备了可生物降解的阳离子肝素-聚乙烯亚胺（HPEI）纳米凝胶，将 pVITRO-IP-10 递送至 SKOV3 人卵巢癌细胞中。通过绿色荧光蛋白的表达谱检测转染效率。通过 RT-PCR 和 Western blot 分析确定 IP-10 的表达。在体外，通过 MTT 测定评估细胞增殖。通过 Hoechst33258/PI 染色和流式细胞术测定评估细胞凋亡。通过人脐静脉内皮细胞（HUVEC）的管形成测定评估对血管生成抑制的影响。此外，建立了 SKOV3 腹腔卵巢癌转移模型，以研究 HPEI+pVITRO-IP-10 复合物在裸鼠中的抗肿瘤活性。在治疗过程中评估肿瘤重量。通过 Ki-67 免疫组织化学染色和 TUNEL 测定评估细胞增殖和凋亡，通过 CD31 免疫组织化学染色和藻酸盐包埋肿瘤细胞测定评估 pVITRO-IP-10 的抗血管生成作用。HPEI 纳米凝胶可有效地将 pVITRO-IP-10 转染到 SKOV3 细胞中。腹腔内给予 HPEI+pVITRO-IP-10 复合物导致卵巢癌的有效生长抑制，其中治疗组的肿瘤重量与空载体对照组相比降低了约 69.92%。同时，与对照组相比，HPEI+pVITRO-IP-10 组观察到细胞增殖减少、肿瘤细胞凋亡增加和血管生成减少。这些结果表明，HPEI 纳米凝胶递送 pVITRO-IP-10 可能在治疗人卵巢癌方面具有价值。
