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Archive > Volume 25 Issue 2 > 2018,25(2):142-147. DOI:10.3872/j.issn.1007-385X.2018.02.006 Prev Next
Basic Research
IL-18 over-expression inhibits proliferation of human colorectal cancer HCT-116 cells
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Abstract:
Objective: To investigate the effects of interleukin-18 over-expression on the in vitro and in vivo proliferation of human colorectal cancer (CRC) HCT-116 cells. Methods: A recombinant lentivirus vector containing human IL-18 gene fragment was constructed.Then theCRC HCT-116 cell line stably expressing human IL-18 (HCT-116/IL-18) was obtained by recombinant lentivirus transfection. In vitro proliferation of HCT-116/IL-18 cells and wild-type HCT-116 cells was determined by CCK-8 method. The expressions of IL-18, Cyclin D1, proliferating cell nuclear antigen (PCNA) and DNA damage repair enzyme (PARP) were detected by Western blotting. HCT-116 and HCT-116/IL-18 cells were inoculated into left and right axillas of Balb/c nude mice, respectively. Then the tumorigenicity and the growth of transplanted tumor were observed. The expressions of IL-18 and PCNA in xenograft tissues were detected by immunohistochemistry analysis. Results: IL-18 gene over-expression in HCT-116 cells could delay the proliferation of HCT-116 cells (P<0.05 or P<0.01). PARP expression was increased significantly and PCNA, Cyclin D1 expression were decreased in HCT-116/IL-18 cells as compared to that of HCT-116 cells (P<0.01).The tumorigenicity of HCT-116/IL-18 cell was significantly decreased in nude mice with a tumor-formation rate of 43%; Compared with control group, HCT-116/IL-18cell line had a longer tumorigensis time,slower growth and smaller tumor volume; moreover, PCNA protein expression was down-regulated in HCT-116/IL-18 xenograft tissue-sas shown by immunohistochemistry analysis (P<0.01). Conclusion: IL-18 over-expression inhibited the growth and proliferation of HCT-116cells both in vitro andin vivo, and the mechanism might be related with IL-18 regulating cell cycle and promotingDNAdamage.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81201765), the Key Scientific Research Projects for Higher Education of Henan Province (No. 15A320063, No.16A310022), the Young Backbone Teachers Training Program of Henan Province (No. 2016GGJS-105)，and the Graduate Student Support of Scientific Research Innovation Projects of Xinxiang Medical University (No. YJSCX201616Y)
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