中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2010年
12期
920-923
,共4页
高国生%翁彭剑%纪礽斌%李德周%李永燕%李红山%丁世雄%胡静
高國生%翁彭劍%紀礽斌%李德週%李永燕%李紅山%丁世雄%鬍靜
고국생%옹팽검%기잉빈%리덕주%리영연%리홍산%정세웅%호정
肝炎病毒,乙型%X基因%SPG21
肝炎病毒,乙型%X基因%SPG21
간염병독,을형%X기인%SPG21
Hepatitis B virus%X gene%SPG21
目的 探讨HBV X基因对痉挛性截瘫蛋白(SPG)21表达的影响.方法 采用RT-PCR和Western blot检测HepG2和HepG2.2.15细胞mRNA和蛋白表达的差异,将带有SPG21基因启动子的报告质粒pGL3-SPG21分别与表达HBV基因组的单个基因的质粒共转染HepG2细胞,测定荧光色素酶的活性,以相对光单元(RUL)表达;RT-PCR和Western blot分别检测SPG21 mRNA和蛋白表达的变化.组间比较采用t检验.结果 HepG2.2.15细胞中SPG21 mRNA和蛋白的表达水平明显高于HepG2细胞,相对表达量(与β-肌动蛋白的灰度比值)为0.36±0.06对比0.21±0.05,P＜0.05.转染pCMV-S、pCMV-E、pCMV-C、pCMV-X、pCMV-P和pCMV-ag2B后的HepG2细胞中,荧光素酶的活性分别为每微克蛋白(86±12)RUL、(75±12)RUL、(69±11)RUL、(875±27)RUL、(104±16)RUL和(67±12)RUL;与转染pCMV-tag2B组细胞相比,转染X基因者荧光素酶活性明显升高(P＜0.01).HBV X基因在mRNA和蛋白水平上调SPG21的表达,这种激活作用随着X基因浓度的增加而增强.结论 HBV X基因能特异性地激活SPG21的表达.
目的 探討HBV X基因對痙攣性截癱蛋白(SPG)21錶達的影響.方法 採用RT-PCR和Western blot檢測HepG2和HepG2.2.15細胞mRNA和蛋白錶達的差異,將帶有SPG21基因啟動子的報告質粒pGL3-SPG21分彆與錶達HBV基因組的單箇基因的質粒共轉染HepG2細胞,測定熒光色素酶的活性,以相對光單元(RUL)錶達;RT-PCR和Western blot分彆檢測SPG21 mRNA和蛋白錶達的變化.組間比較採用t檢驗.結果 HepG2.2.15細胞中SPG21 mRNA和蛋白的錶達水平明顯高于HepG2細胞,相對錶達量(與β-肌動蛋白的灰度比值)為0.36±0.06對比0.21±0.05,P＜0.05.轉染pCMV-S、pCMV-E、pCMV-C、pCMV-X、pCMV-P和pCMV-ag2B後的HepG2細胞中,熒光素酶的活性分彆為每微剋蛋白(86±12)RUL、(75±12)RUL、(69±11)RUL、(875±27)RUL、(104±16)RUL和(67±12)RUL;與轉染pCMV-tag2B組細胞相比,轉染X基因者熒光素酶活性明顯升高(P＜0.01).HBV X基因在mRNA和蛋白水平上調SPG21的錶達,這種激活作用隨著X基因濃度的增加而增彊.結論 HBV X基因能特異性地激活SPG21的錶達.
목적 탐토HBV X기인대경련성절탄단백(SPG)21표체적영향.방법 채용RT-PCR화Western blot검측HepG2화HepG2.2.15세포mRNA화단백표체적차이,장대유SPG21기인계동자적보고질립pGL3-SPG21분별여표체HBV기인조적단개기인적질립공전염HepG2세포,측정형광색소매적활성,이상대광단원(RUL)표체;RT-PCR화Western blot분별검측SPG21 mRNA화단백표체적변화.조간비교채용t검험.결과 HepG2.2.15세포중SPG21 mRNA화단백적표체수평명현고우HepG2세포,상대표체량(여β-기동단백적회도비치)위0.36±0.06대비0.21±0.05,P＜0.05.전염pCMV-S、pCMV-E、pCMV-C、pCMV-X、pCMV-P화pCMV-ag2B후적HepG2세포중,형광소매적활성분별위매미극단백(86±12)RUL、(75±12)RUL、(69±11)RUL、(875±27)RUL、(104±16)RUL화(67±12)RUL;여전염pCMV-tag2B조세포상비,전염X기인자형광소매활성명현승고(P＜0.01).HBV X기인재mRNA화단백수평상조SPG21적표체,저충격활작용수착X기인농도적증가이증강.결론 HBV X기인능특이성지격활SPG21적표체.
Objective To investigate the effect of hepatitis B virus(HBV) X gene on the expression of SPG21. Methods The expressions of SPG21 mRNA and protein in HepG2 and HepG2.2.15 cells were tested by RT-PCR and western blot. HepG2 cells were co-transfected with reporter plasmid pGL3-SPG21 and plasmids carrying individual genes of HBV, the luciferase activity was measured and the expressions of SPG21 were detected by RT-PCR and western blot. Results The expressions of SPG21 mRNA and protein were higher in HepG2.2.15 cells than in HepG2 cells (0.36 ± 0.06 vs 0.21 ± 0.05, P ＜ 0.05). The activity of SPG21 in HepG2 cells transfected with pCMV-X was higher (875 ± 27 vs 67 ± 12, P ＜ 0.01) as compared to blank control group (transfected with pCMV-tag2B). HBV X gene enhanced SPG21 gene promoter activity,SPG21 mRNA expression and SPG21 protein production in HepG2 cells in a dose-dependent manner. Conclusions HBV X gene can specially activate SPG21 expression.
