CAJ | 학술논문

目的探讨p16表达与胃癌细胞的增殖能力、胃癌的临床特征及胃癌患者预后的关系.方法采用逆转录聚合酶链反应(RT-PCR)、免疫组化和Westem blot方法,检测胃癌细胞系SGC-7901、MKN45、MKN28和65例胃癌组织标本中p16 mRNA和蛋白的表达.以AdCMV-p16感染胃癌SGC-7901细胞,采用四甲基偶氮唑蓝(MTT)法测定感染前后SGC-7901细胞存活率的变化,采用流式细胞术检测感染前后SGC-7901细胞细胞周期的变化.比较不同p16表达状态胃癌患者的术后生存情况.结果胃癌细胞系MKN45呈p16弱阳性表达,SGC-7901和MKN28细胞系呈阴性表达.以20PFU/细胞感染腺病毒AdCMV-p16时,原本p16阴性表达的SGC-7901细胞中p16表达转为阳性.转染p16的SGC-7901细胞存活率下降,G1期细胞比例上升,S期细胞比例下降.65例胃癌组织中,p16阳性率为32.3％,明显低于正常胃黏膜组织(81.5％),且p16表达与胃癌的分化程度、淋巴结转移有关.p16阴性组的胃癌患者无瘤生存期明显缩短.结论 p16基因对胃癌细胞增殖的调控至关重要,p16的失活赋予了胃癌细胞高增殖、转移活性,并导致了胃癌患者的无瘤生存期缩短.
목적 탐토p16표체여위암세포적증식능력、위암적림상특정급위암환자예후적관계.방법 채용역전록취합매련반응(RT-PCR)、면역조화화Westem blot방법,검측위암세포계SGC-7901、MKN45、MKN28화65례위암조직표본중p16 mRNA화단백적표체.이AdCMV-p16감염위암SGC-7901세포,채용사갑기우담서람(MTT)법측정감염전후SGC-7901세포존활솔적변화,채용류식세포술검측감염전후SGC-7901세포세포주기적변화.비교불동p16표체상태위암환자적술후생존정황.결과 위암세포계MKN45정p16약양성표체,SGC-7901화MKN28세포계정음성표체.이20PFU/세포감염선병독AdCMV-p16시,원본p16음성표체적SGC-7901세포중p16표체전위양성.전염p16적SGC-7901세포존활솔하강,G1기세포비례상승,S기세포비례하강.65례위암조직중,p16양성솔위32.3％,명현저우정상위점막조직(81.5％),차p16표체여위암적분화정도、림파결전이유관.p16음성조적위암환자무류생존기명현축단.결론 p16기인대위암세포증식적조공지관중요,p16적실활부여료위암세포고증식、전이활성,병도치료위암환자적무류생존기축단.
Objective To investigate the relationship between p16 expression and cell proliferation and prognosis in gastric cancer patients.Methods Gastric cancer cell lines SGC-7901,MKN45,MKN28,human embryonic kidney cell line HEK293,human fibroblast cell line MRC-5,and surgical specimens of gastric carcinoma and adjacent normal gastric mucosa from 65 patients were included in this study.RT-PCR,MTT and FCM assays were used to detect p16 expression in gastric cancer cell lines and surgical specimens of gastric cancer.MTT assay was used to determine cancer cell viability and FCM to detect cell cycle.Kaplan-Meier survival curve and Log-Rank statistics were used to analyze the relationship between p16expression and survival of petients with gastric cancer.Results Gastric cancer cell lines were mostly negative for p16 expression,and p16 was re-expressed after the cells transfeeted with p16 gene by adenovirus AdCMV-p16.p16 re-expression resulted in the decrease of cancer cell viability and cancer cell cycle arrest with increased G1 phase and decreased S phase.p16 expression in cancer specimens was 32.3％ (21/65),significantly lower than the 81.5％ (53/65) in normal mucosa (x2 =32.124,P ＜ 0.001 ).The disease-free survival was significantly shorter in p16-negative patients than that in p16-positive patients (P ＜0.01 ),but not the overall survival ( P ＞ 0.05 ).p16 expression was significantly correlated with differentiation and lymph node metastasis,but not significantly correlated with sex,age,tumor size or invasion depth of the gastric cancer.Conclusions p16 gene is important for cancer cell proliferation.The inactivation gives cancer cells a high activity for proliferation and metastasis,and then influences the disease-free survival of gastric cancer patients.