Adv-mIL-12转染KC对人结直肠癌细胞生长影响的实验研究

目的 探讨Adv-mIL-12转染KC对人结直肠癌细胞的生长抑制作用.方法 腺病毒Adv-mIL-12体外转染KC,EHSA法检测其IL-12的表达;将KC,Kc-Adr-EGFP和KC-Adv-mIL-12与LoVo人结直肠癌细胞共培养,MTT和ELISA法检测其对LoVo细胞的生长抑制率和VEGF表达量的影响;FCM检测不同KC组的上清液诱导LoVo细胞凋亡的差异.结果 KC-Adv-mIL-12中IL-12浓度明显提高[(564.29±31.24)Pg/mL 24 h];MTT,ELISA和FCM等证实KC-Adv-mIL-12对LoVo细胞的抑制作用较另2个对照组明显(P<0.05).LoVo细胞的凋亡率显著增加.结论 mIL-12基因转染KC能有效抑制人结直肠癌细胞的生长及转移.

Experimental study of Kupffer cell transfected with a recombinant adenoviral vector expressing interleukin 12 on proliferation of LoVo human colorectal cancer cells

Objective:To explore the effect of Kupffer Cells (KC) transfected with a recombinant adenoviral vector expressing interleukin 12 (IL-12) on human colorectal cancer LoVo cells. Methods:ELISA method was used to detect quantitatively the level of IL-12 of Kupffer cells transfected with Adv-mIL-12. KC,KC-adv-EGFP and KC-Adr-mlL-12 with LoVo cells were cultured in different plates. The cells viability and the supernatants′ VEGF were measured by MTT assay and VEGF ELISA kit；different supernatants groups of LoVo cells were harvested and differences of apoptosis were analyzed using FCM. Results:A high level of secretion of IL-12 was detected in the adv-mIL-12-transfected KC［(564.29±31.24)pg/mL 24 h］. The MTT assay and ELISA test showed that the inhibition ratio of LoVo cells was higher in the experimental group compared with 2 control groups（P＜0.05）；the FCM showed that more apoptosis cells were detected in the the experimental group(21%).Conclusions:KC-Adv-mIL-12 can inhibit the proliferation and metastasis of LoVo cells.