siRNA 沉默肾上腺髓质素基因对 A375黑素瘤细胞表达 VEGF IL－10的影响

摘要

Objective: To assess the impact of small interference RNA-induced silencing of ad-renomedullin gene on the expression of VEGF, IL-10 in malignant melanoma cells.Method: A375 cells were classified into 3 groups to be transfected with the selected siRNA( test group) , non-specific sequences (negative control group), or remain untransfected(blank control group).Immunocytochemical staining and ELISA was performed to detect the expressions of VEGF and IL-10 in each group.Result:The expression of VEGF was 192.246 ±17.330 pg/mL in the control group,187.831 ±14.450 pg/mL in the negative control group and 96.218±10.078 pg/mL in ADMsiRNA transfected group.The expression of IL-10 was 108.822±7. 55 pg/mL in the control group, 103.250±4.785 pg/mL in the negative control group and 37.024±5.770 pg/mL in ADMsiRNA transfected group.The differences were statistically significant.The results of immunohis-tochemistry are the same of ELISA.Conclusion:siRNA silencing adrenomedullin gene can inhibit the expres-sion of VEGF and IL-10 in A375 melanoma cell.%目的：研究siRNA沉默肾上腺髓质素基因对A375黑素瘤细胞表达VEGF、IL－10的影响。方法：采用人黑素瘤细胞株A375，分为采用正常未转染组细胞做为空白对照，人工合成的非特异序列转染A375细胞为阴性对照，人工合成的肾上腺髓质素（ ADM）靶向小分子干扰RNA转染A375细胞为抑制ADM基因表达实验组。采用酶联免疫吸附试验（ ELISA）方法和免疫细胞化学染色方法检测各组细胞中VEGF、IL－10的表达水平。结果：空白对照组、阴性对照组、ADM siRNA 转染组ELISA法测定VEGF的表达量分别为192．246±17．330、187．831±14．450、96．218±10．078 pg／mL；IL－10的表达量分别为108．822±7．55、103．250±4．785、37．024±5．770pg／mL，组间比较差异均有统计学意义（ P＜0．01或P＜0．05）。免疫组化法测定显示结果同ELISA结果。结论：siRNA沉默ADM基因可下调A375黑素瘤细胞免疫抑制分子VEGF、IL－10的表达。