中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
9期
1422-1424
,共3页
扈玉华%吴建梁%孟宪兵%陈泽
扈玉華%吳建樑%孟憲兵%陳澤
호옥화%오건량%맹헌병%진택
c-myc基因%RNA干扰%血管内皮生长因子%胶质瘤
c-myc基因%RNA榦擾%血管內皮生長因子%膠質瘤
c-myc기인%RNA간우%혈관내피생장인자%효질류
c-myc gene%RNA interference%Vascular endothelial growth factor%Glioma
目的 观察c-myc癌基因在体外对胶质瘤细胞血管内皮生子因子(VEGF)表达的影响。方法 构建以c-myc基因为靶向的shRNA干扰质粒pCMYC,同时构建pHK质粒作为阴性对照,采用逆转录-聚合酶链反应(RT-PCR)方法检测各组细胞中c-myc及VEGF mRNA水平的表达变化,采用免疫细胞化学法及Western blot法分别检测3组细胞中c-myc及VEGF蛋白的表达。结果 (1) IN500Δ-pCMYC细胞组c-myc与VEGF mRNA表达降低(0.273±0.028,0.443 ±0.048),与IN500Δ细胞组(1.185±0.145,1.116±0.072)及IN500Δ-pHK细胞组(1.098±0.128、1.208±0.133)比较差异均有统计学意义(P＜0.01)。(2)免疫细胞化学染色结果显示,IN500Δ-pCMYC细胞组c-myc与VEGF阳性细胞表达率均显著降低,为(24.52 ±4.39)％及(23.52±3.44)％,与IN500Δ及IN500Δ-pHK细胞组比较差异有统计学意义(P＜0.05)。(3)Western blot法检测结果显示IN500Δ-pCMYC细胞中c-myc与VEGF的蛋白表达降低(0.221±0.041、0.284±0.025),与IN500Δ细胞组(1.821±0.191、1.531±0.213)及1N500Δ-pHK细胞组(1.650±0.110、1.820±0.122)比较差异均有统计学意义(P＜0.01)。结论 沉默c-myc基因能够抑制胶质瘤中VEGF的表达,从而降低肿瘤组织内的血管生成。
目的 觀察c-myc癌基因在體外對膠質瘤細胞血管內皮生子因子(VEGF)錶達的影響。方法 構建以c-myc基因為靶嚮的shRNA榦擾質粒pCMYC,同時構建pHK質粒作為陰性對照,採用逆轉錄-聚閤酶鏈反應(RT-PCR)方法檢測各組細胞中c-myc及VEGF mRNA水平的錶達變化,採用免疫細胞化學法及Western blot法分彆檢測3組細胞中c-myc及VEGF蛋白的錶達。結果 (1) IN500Δ-pCMYC細胞組c-myc與VEGF mRNA錶達降低(0.273±0.028,0.443 ±0.048),與IN500Δ細胞組(1.185±0.145,1.116±0.072)及IN500Δ-pHK細胞組(1.098±0.128、1.208±0.133)比較差異均有統計學意義(P＜0.01)。(2)免疫細胞化學染色結果顯示,IN500Δ-pCMYC細胞組c-myc與VEGF暘性細胞錶達率均顯著降低,為(24.52 ±4.39)％及(23.52±3.44)％,與IN500Δ及IN500Δ-pHK細胞組比較差異有統計學意義(P＜0.05)。(3)Western blot法檢測結果顯示IN500Δ-pCMYC細胞中c-myc與VEGF的蛋白錶達降低(0.221±0.041、0.284±0.025),與IN500Δ細胞組(1.821±0.191、1.531±0.213)及1N500Δ-pHK細胞組(1.650±0.110、1.820±0.122)比較差異均有統計學意義(P＜0.01)。結論 沉默c-myc基因能夠抑製膠質瘤中VEGF的錶達,從而降低腫瘤組織內的血管生成。
목적 관찰c-myc암기인재체외대효질류세포혈관내피생자인자(VEGF)표체적영향。방법 구건이c-myc기인위파향적shRNA간우질립pCMYC,동시구건pHK질립작위음성대조,채용역전록-취합매련반응(RT-PCR)방법검측각조세포중c-myc급VEGF mRNA수평적표체변화,채용면역세포화학법급Western blot법분별검측3조세포중c-myc급VEGF단백적표체。결과 (1) IN500Δ-pCMYC세포조c-myc여VEGF mRNA표체강저(0.273±0.028,0.443 ±0.048),여IN500Δ세포조(1.185±0.145,1.116±0.072)급IN500Δ-pHK세포조(1.098±0.128、1.208±0.133)비교차이균유통계학의의(P＜0.01)。(2)면역세포화학염색결과현시,IN500Δ-pCMYC세포조c-myc여VEGF양성세포표체솔균현저강저,위(24.52 ±4.39)％급(23.52±3.44)％,여IN500Δ급IN500Δ-pHK세포조비교차이유통계학의의(P＜0.05)。(3)Western blot법검측결과현시IN500Δ-pCMYC세포중c-myc여VEGF적단백표체강저(0.221±0.041、0.284±0.025),여IN500Δ세포조(1.821±0.191、1.531±0.213)급1N500Δ-pHK세포조(1.650±0.110、1.820±0.122)비교차이균유통계학의의(P＜0.01)。결론 침묵c-myc기인능구억제효질류중VEGF적표체,종이강저종류조직내적혈관생성。
Objective To explore the expression of vascular endothelial growth factor (VEGF) in glioblastoma cells after silencing the oncogene c-myc through RNA interference (RNAi). Methods The shRNA interference plasmid named pCMYC targeting c-myc gene and the pHK plasmid as negative control were constructed. The expression of c-myc and VEGF mRNA and protein was detected by using reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry and Western blotting among different groups. Results ( 1 ) The mRNA expression of c-myc and VEGF in IN500Δ-pCMYC cells (0. 273 ±0. 028, 0. 443 ±0. 048) was lower than that of IN500Δ ( 1. 185 ±0. 145, 1. 116 ±0. 072) and IN500Δ-pHK cells ( 1. 098 t0. 128, 1. 208 ±0. 133) (P ＜0. 01 ) ; (2) Immunocytochemical staining revealed that the expression of c-myc and VEGF protein in IN500Δ-pCMYC cells was lower than in IN500Δ cells and IN500Δ-pHK cells (P＜0. 05); (3) Western blotting indicated that the expression of c-myc and VEGF protein in IN500Δ-pCMYC cells (0. 221 ± 0. 041, 0. 284 ± 0. 025 ) was lower than that in IN500Δ cells (1.821±0.191, 1.531 ±0.213) and IN500A-pHK cells (1.650±0.110, 1.820±0.122) (P＜0. 01 ). Conclusion Sliencing the expression of c-myc oncogene could effectively inhibit the expression of VEGF.
