损伤相关模式分子对人肝癌HepG2细胞增殖能力的影响

Effect of DAMPs on proliferation of human liver cancer HepG2 cells

目的 观察在DAMPs诱导下人肝癌HepG2细胞增殖能力的变化.方法 将HepG2细胞分为对照组和实验组（10、20、40、80 μl DAMPs处理的HepG2细胞）;MTT比色法检测HepG2细胞的增殖能力;实时荧光定量PCR法检测IL-6 mRNA表达的变化;Western Blot法检测HepG2细胞IL-6蛋白的表达情况.结果 HepG2细胞随着DAMPs剂量的递增及时间的延长增殖能力逐渐增强,呈现明显的量效-时效关系,在剂量40 μl,作用时间36 h时细胞增殖能力达到最强,差异有统计学意义（P〈0.01）;选取作用时间为36 h,随着DAMPs剂量的递增,实时定量PCR法检测到HepG2细胞IL-6 mRNA分别为95.55±4.47,171.80±6.60,453.30±14.47,610.59±12.70,441.04±18.91,差异有统计学意义（P〈0.01）;Western Blot法检测HepG2细胞IL-6蛋白的表达分别为1.47、2.07、2.74、3.44、3.00,差异有统计学意义（P〈0.01）.结论 DAMPs在一定剂量及时间内促进人肝癌HepG2细胞增殖,并且呈现明显的量效-时效关系.

Objective To study the changes in DAMPs-induced proliferation of liver cancer HepG2 ceils. Methods HepG2 cells were divided into control group, and DAMPs-treated groups （10, 20, 40 and 80 L）. MTT assay was applied to investigate the proliferation of HepG2 cells. The expression of IL-6 mRNA in HepG2 cells was detected by using quantitative RT-PCR. Results MTT result demonstrated that the proliferation of HepG2 cells had a dose- and time-dependent with DAMPs. The proliferation of HepG2 cells reached to peak when the dose of DAMPs was 40 L for 36 h of culture （P〈0. 01）. Quantitative RT-PCR revealed that the expression of IL-6 mRNA was 95.55 ±4. 47, 171.80 ± 6.60, 453.30 ± 14.47, 610. 59 ± 12. 70 and 441.04± 18. 91 respectively when HepG2 cells were exposed to DAMPs （10, 20, 40 and 80 L） at 36 h （P〈0. 01）, while thelL-6 pro- tein level in HepG2 cells was 1.47, 2. 07, 2.74, 3.44, 3.00 respectively （P〈0. 01）. Conclusion DAMPs may promote the proliferation of HepG2 cells in a dose-and time-dependent manner.