中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
30期
2140-2143
,共4页
刘香娟%李福年%姜丹丹%王新刚%刘相萍%张佃良%孟春晖
劉香娟%李福年%薑丹丹%王新剛%劉相萍%張佃良%孟春暉
류향연%리복년%강단단%왕신강%류상평%장전량%맹춘휘
基因,myc%基因表达调控%细胞分裂%细胞凋亡
基因,myc%基因錶達調控%細胞分裂%細胞凋亡
기인,myc%기인표체조공%세포분렬%세포조망
Genes,myc%Gene expression regulation%Cell division%Apoptosis
目的 了解非p53依赖通路突变型与野生型c-myc基因对p14ARF基因表达调控及细胞凋亡的诱导功能.方法 分别用携带突变型与野生型c -myc基因的慢病毒载体感染p53缺失型乳腺癌HCC1937细胞并得到二者过表达的稳定细胞株,未感染组及感染不携带c-myc基因的慢病毒细胞作空白对照和感染对照,采用反转录( RT) -PCR、Western印迹分别检测突变型与野生型c-myc及p14ARF基因mRNA和蛋白表达,采用MTT、原位末端标记(TUNEL)检测突变型与野生型c-myc基因过表达乳腺癌HCC1937细胞增殖与凋亡情况.结果 RT-PCR和Western印迹均显示突变型组与野生型组c-myc基因mRNA和蛋白过表达,突变型与野生型组c-myc蛋白[c-myc/β-肌动蛋白(actin)]表达量分别为0.560 ±0.010、0.651 ±0.010,与两对照组相比差异具有统计学意义(P＜0.05)；野生型组p14ARF蛋白(p14ARF/β-actin)表达量为0.382±0.013,与两对照组相比差异均有统计学意义(均P＜0.05)；突变型组p14ARF蛋白(p14ARF/β-actin)表达量为0.154 ±0.011,与两对照组相比,差异均无统计学意义(均P＞0.05).突变型组细胞增殖活性显著高于野生型组(P＜0.05),突变型组与两对照组细胞凋亡率比较差异均无统计学意义(均P ＞0.05),野生型组细胞凋亡率显著高于突变型组与两对照组(空白对照组为3.8％±0.5％,感染对照组为3.8％±0.4％,突变型组为3.2％±0.4％,野生型组为7.1％±0.7％)(均P＜0.05).结论 非p53依赖性通路中,野生型c-myc基因过表达能明显上调p14ARF基因表达、诱导细胞凋亡,其促进增殖功能与诱导凋亡功能保持平衡,而突变后此作用丧失,致瘤性增加.
目的 瞭解非p53依賴通路突變型與野生型c-myc基因對p14ARF基因錶達調控及細胞凋亡的誘導功能.方法 分彆用攜帶突變型與野生型c -myc基因的慢病毒載體感染p53缺失型乳腺癌HCC1937細胞併得到二者過錶達的穩定細胞株,未感染組及感染不攜帶c-myc基因的慢病毒細胞作空白對照和感染對照,採用反轉錄( RT) -PCR、Western印跡分彆檢測突變型與野生型c-myc及p14ARF基因mRNA和蛋白錶達,採用MTT、原位末耑標記(TUNEL)檢測突變型與野生型c-myc基因過錶達乳腺癌HCC1937細胞增殖與凋亡情況.結果 RT-PCR和Western印跡均顯示突變型組與野生型組c-myc基因mRNA和蛋白過錶達,突變型與野生型組c-myc蛋白[c-myc/β-肌動蛋白(actin)]錶達量分彆為0.560 ±0.010、0.651 ±0.010,與兩對照組相比差異具有統計學意義(P＜0.05)；野生型組p14ARF蛋白(p14ARF/β-actin)錶達量為0.382±0.013,與兩對照組相比差異均有統計學意義(均P＜0.05)；突變型組p14ARF蛋白(p14ARF/β-actin)錶達量為0.154 ±0.011,與兩對照組相比,差異均無統計學意義(均P＞0.05).突變型組細胞增殖活性顯著高于野生型組(P＜0.05),突變型組與兩對照組細胞凋亡率比較差異均無統計學意義(均P ＞0.05),野生型組細胞凋亡率顯著高于突變型組與兩對照組(空白對照組為3.8％±0.5％,感染對照組為3.8％±0.4％,突變型組為3.2％±0.4％,野生型組為7.1％±0.7％)(均P＜0.05).結論 非p53依賴性通路中,野生型c-myc基因過錶達能明顯上調p14ARF基因錶達、誘導細胞凋亡,其促進增殖功能與誘導凋亡功能保持平衡,而突變後此作用喪失,緻瘤性增加.
목적 료해비p53의뢰통로돌변형여야생형c-myc기인대p14ARF기인표체조공급세포조망적유도공능.방법 분별용휴대돌변형여야생형c -myc기인적만병독재체감염p53결실형유선암HCC1937세포병득도이자과표체적은정세포주,미감염조급감염불휴대c-myc기인적만병독세포작공백대조화감염대조,채용반전록( RT) -PCR、Western인적분별검측돌변형여야생형c-myc급p14ARF기인mRNA화단백표체,채용MTT、원위말단표기(TUNEL)검측돌변형여야생형c-myc기인과표체유선암HCC1937세포증식여조망정황.결과 RT-PCR화Western인적균현시돌변형조여야생형조c-myc기인mRNA화단백과표체,돌변형여야생형조c-myc단백[c-myc/β-기동단백(actin)]표체량분별위0.560 ±0.010、0.651 ±0.010,여량대조조상비차이구유통계학의의(P＜0.05)；야생형조p14ARF단백(p14ARF/β-actin)표체량위0.382±0.013,여량대조조상비차이균유통계학의의(균P＜0.05)；돌변형조p14ARF단백(p14ARF/β-actin)표체량위0.154 ±0.011,여량대조조상비,차이균무통계학의의(균P＞0.05).돌변형조세포증식활성현저고우야생형조(P＜0.05),돌변형조여량대조조세포조망솔비교차이균무통계학의의(균P ＞0.05),야생형조세포조망솔현저고우돌변형조여량대조조(공백대조조위3.8％±0.5％,감염대조조위3.8％±0.4％,돌변형조위3.2％±0.4％,야생형조위7.1％±0.7％)(균P＜0.05).결론 비p53의뢰성통로중,야생형c-myc기인과표체능명현상조p14ARF기인표체、유도세포조망,기촉진증식공능여유도조망공능보지평형,이돌변후차작용상실,치류성증가.
Objective To explore the regulation of p14ARF expression and induction of cell apoptosis with the mutant and wild-type c-myc genes in a p53-independent pathway of signal transduction.Methods The mutant and wild-type c-myc genes were transfected by lentivirus into HCC1937 to form the stable over-expression cell lines.Uninfected cells and lentivirus-infected ones carrying no c-myc gene acted as blank and infection controls respectively. And c-myc and p14ARF mRNA and protein,proliferation and apoptosis in HCC1937 with mutant and Mild-type c-myc were detected by reverse transcription (RT)-PCR,Western blotting,thiazolyl blue tetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling (TUNEL) respectively.Results After the lentivirus-mediated gene transfer,c-myc mRNA and protein expression increased in the mutant and wild-type groups.p14ARF mRNA and protein increased in the wild-type group and the mutant group and there were significant difference between them with blank and infection controls (mutant groups:0.560 ± 0.010,0.154 ± 0.011,wild-type groups:0.651 ±0.010,0.382 ± 0.013,both P ＜ 0.05 ).The group of mutant and wild-type c-myc could promote the proliferation of cell growth. And c-myc was more effective to induce apoptosis in the wild-type group as compared with the mutant group (7.1％ ± 0.7％ vs 3.2％ ± 0.4％,P ＜ 0.05 ).Conclusions In a p53-independent pathway,the over-expression of wild-type c-myc obviously up-regulates the expression of p14ARF.And cell apoptosis may be induced through the regulation of p14ARF-related gene,keep balance of proliferative promotion and apoptosis induction. When there is a loss-of-function of mutant c-myc,tumorigenicity increases via a disturbed balance of proliferative promotion and apoptosis induction.
