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
1419-1421
,共3页
杨洪宽%毛峰%王宝峰%万峰%郭东生%雷霆
楊洪寬%毛峰%王寶峰%萬峰%郭東生%雷霆
양홍관%모봉%왕보봉%만봉%곽동생%뢰정
神经胶质瘤%LRIG3%增殖细胞核抗原%Ki-67
神經膠質瘤%LRIG3%增殖細胞覈抗原%Ki-67
신경효질류%LRIG3%증식세포핵항원%Ki-67
Glioma%LRIG3%Proliferating cell nuclear antigen%Ki-67
目的 探讨过表达LRIG3基因对神经胶质瘤细胞株U251与U87增殖及增殖细胞核抗原(PCNA)和Ki-67表达的影响及其机制。方法 用携带LRIG3过表达特异性序列和只含空白载体的质粒用慢病毒法感染胶质瘤细胞株U251与U87,筛选稳定株,分别分成实验组和对照组,通过逆转录-聚合酶链反应(RT-PCR)和Western blot法检测各组细胞LRIG3表达的变化,应用噻唑蓝(MTT)比色法检测病毒感染后对胶质瘤细胞株U251与U87增殖的影响,用免疫组织化学链霉亲和素生物素复合物(SABC)法分别检测各组细胞中PCNA和Ki-67的表达差异。结果 LRIG3过表达组细胞中LRIG3 mRNA水平与对照组比较分别升高67.6％( U251)和79.9％ (U87),LRIG3蛋白表达水平分别升高62.3％(U87)和91.0％( U251)。MTT法结果显示两实验组细胞增殖率均低于相应对照组。U251细胞对照组PCNA阳性率为(47.81 ±4.67)％,实验组为(27.49±3.17)％,U87细胞对照组PCNA阳性率为(55.50±4.01)％,实验组为(33.60±4.82)％,差异均有统计学意义(P＜0.05)。U251细胞对照组中Ki-67的阳性率为(48.50±6.11)％,实验组为(24.30±3.76)％,U87细胞对照组Ki-67阳性率为(55.20±4.19)％,实验组为(23.50±4.60)％,差异均有统计学意义(P＜0.0l)。结论 LRIG3基因过表达可减少胶质瘤细胞的增殖。
目的 探討過錶達LRIG3基因對神經膠質瘤細胞株U251與U87增殖及增殖細胞覈抗原(PCNA)和Ki-67錶達的影響及其機製。方法 用攜帶LRIG3過錶達特異性序列和隻含空白載體的質粒用慢病毒法感染膠質瘤細胞株U251與U87,篩選穩定株,分彆分成實驗組和對照組,通過逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot法檢測各組細胞LRIG3錶達的變化,應用噻唑藍(MTT)比色法檢測病毒感染後對膠質瘤細胞株U251與U87增殖的影響,用免疫組織化學鏈黴親和素生物素複閤物(SABC)法分彆檢測各組細胞中PCNA和Ki-67的錶達差異。結果 LRIG3過錶達組細胞中LRIG3 mRNA水平與對照組比較分彆升高67.6％( U251)和79.9％ (U87),LRIG3蛋白錶達水平分彆升高62.3％(U87)和91.0％( U251)。MTT法結果顯示兩實驗組細胞增殖率均低于相應對照組。U251細胞對照組PCNA暘性率為(47.81 ±4.67)％,實驗組為(27.49±3.17)％,U87細胞對照組PCNA暘性率為(55.50±4.01)％,實驗組為(33.60±4.82)％,差異均有統計學意義(P＜0.05)。U251細胞對照組中Ki-67的暘性率為(48.50±6.11)％,實驗組為(24.30±3.76)％,U87細胞對照組Ki-67暘性率為(55.20±4.19)％,實驗組為(23.50±4.60)％,差異均有統計學意義(P＜0.0l)。結論 LRIG3基因過錶達可減少膠質瘤細胞的增殖。
목적 탐토과표체LRIG3기인대신경효질류세포주U251여U87증식급증식세포핵항원(PCNA)화Ki-67표체적영향급기궤제。방법 용휴대LRIG3과표체특이성서렬화지함공백재체적질립용만병독법감염효질류세포주U251여U87,사선은정주,분별분성실험조화대조조,통과역전록-취합매련반응(RT-PCR)화Western blot법검측각조세포LRIG3표체적변화,응용새서람(MTT)비색법검측병독감염후대효질류세포주U251여U87증식적영향,용면역조직화학련매친화소생물소복합물(SABC)법분별검측각조세포중PCNA화Ki-67적표체차이。결과 LRIG3과표체조세포중LRIG3 mRNA수평여대조조비교분별승고67.6％( U251)화79.9％ (U87),LRIG3단백표체수평분별승고62.3％(U87)화91.0％( U251)。MTT법결과현시량실험조세포증식솔균저우상응대조조。U251세포대조조PCNA양성솔위(47.81 ±4.67)％,실험조위(27.49±3.17)％,U87세포대조조PCNA양성솔위(55.50±4.01)％,실험조위(33.60±4.82)％,차이균유통계학의의(P＜0.05)。U251세포대조조중Ki-67적양성솔위(48.50±6.11)％,실험조위(24.30±3.76)％,U87세포대조조Ki-67양성솔위(55.20±4.19)％,실험조위(23.50±4.60)％,차이균유통계학의의(P＜0.0l)。결론 LRIG3기인과표체가감소효질류세포적증식。
Objective To explore the effect of overexpression of LRIG3 gene on proliferation of glioma cells and expression of proliferating cell nuclear antigen (PCNA) and Ki-67, and the possible mechanisms. Methods The plasmid containing LRIG3 gene and control was transduced into glioma U251 and U87 cells respectively. The mRNA and protein levels of LRIG3 were detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Cell proliferation was examined by methyl thiazol tetrazolium (MTT) assay. The expression of PCNA and Ki-67 was detected by strept avidinbiotin complex (SABC). Results Compared with control cells, the mRNA levels of LRIG3 were raised by 67.6％ and 79.9％ in LRIG3 cells of U87 and U251, respectively, and the protein levels were increased by 62. 3％ (U87) and 91.0％ ( U251 ) respectively. Overexpression of LRIG3 resulted in the reduction of cell proliferation. The positive rate of PCNA was significantly lower in LRIG3 cells than in control cells [U87: (33.60±4.82)％ vs (55.50±4.01)％; U251: (27.49±3.17)％ vs (47.81 ±4.67)％ (P＜0. 05)]. The positive rate of Ki-67 was also significantly decreased in LRIG3 transduced cells as compared with control cells[U87 : (23. 50 ±4. 60)％ vs (55.20 ±4. 19)％ ; U251 : (24. 30 ± 3. 76) ％ vs (48. 50 ± 6. 11 ) ％ ( P ＜ 0. 01 )]. Conclusion Up-regulating LRIG3 gene expression can reduce the proliferation of glioma U87 and U251 cells.
