河北医学
河北醫學
하북의학
HEBEI MEDICINE
2015年
7期
1096-1099
,共4页
高阳%胡晓梅%杨宁%孙宇%梁官钊%韩晨珠%段昕所
高暘%鬍曉梅%楊寧%孫宇%樑官釗%韓晨珠%段昕所
고양%호효매%양저%손우%량관쇠%한신주%단흔소
小分子干扰%肾上腺髓质素%黑素瘤%血管内皮生长因子%白细胞介素-10
小分子榦擾%腎上腺髓質素%黑素瘤%血管內皮生長因子%白細胞介素-10
소분자간우%신상선수질소%흑소류%혈관내피생장인자%백세포개소-10
small interfering%Adrenomedullin%Melanoma%VEGF%IL-10
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。
目的:研究siRNA沉默腎上腺髓質素基因對A375黑素瘤細胞錶達VEGF、IL-10的影響。方法:採用人黑素瘤細胞株A375,分為採用正常未轉染組細胞做為空白對照,人工閤成的非特異序列轉染A375細胞為陰性對照,人工閤成的腎上腺髓質素( ADM)靶嚮小分子榦擾RNA轉染A375細胞為抑製ADM基因錶達實驗組。採用酶聯免疫吸附試驗( ELISA)方法和免疫細胞化學染色方法檢測各組細胞中VEGF、IL-10的錶達水平。結果:空白對照組、陰性對照組、ADM siRNA 轉染組ELISA法測定VEGF的錶達量分彆為192.246±17.330、187.831±14.450、96.218±10.078 pg/mL;IL-10的錶達量分彆為108.822±7.55、103.250±4.785、37.024±5.770pg/mL,組間比較差異均有統計學意義( P<0.01或P<0.05)。免疫組化法測定顯示結果同ELISA結果。結論:siRNA沉默ADM基因可下調A375黑素瘤細胞免疫抑製分子VEGF、IL-10的錶達。
목적:연구siRNA침묵신상선수질소기인대A375흑소류세포표체VEGF、IL-10적영향。방법:채용인흑소류세포주A375,분위채용정상미전염조세포주위공백대조,인공합성적비특이서렬전염A375세포위음성대조,인공합성적신상선수질소( ADM)파향소분자간우RNA전염A375세포위억제ADM기인표체실험조。채용매련면역흡부시험( ELISA)방법화면역세포화학염색방법검측각조세포중VEGF、IL-10적표체수평。결과:공백대조조、음성대조조、ADM siRNA 전염조ELISA법측정VEGF적표체량분별위192.246±17.330、187.831±14.450、96.218±10.078 pg/mL;IL-10적표체량분별위108.822±7.55、103.250±4.785、37.024±5.770pg/mL,조간비교차이균유통계학의의( P<0.01혹P<0.05)。면역조화법측정현시결과동ELISA결과。결론:siRNA침묵ADM기인가하조A375흑소류세포면역억제분자VEGF、IL-10적표체。
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.