中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2014年
9期
675-681
,共7页
盛伟伟%董明%周建平%李新%柳青峰%董齐%李丰
盛偉偉%董明%週建平%李新%柳青峰%董齊%李豐
성위위%동명%주건평%리신%류청봉%동제%리봉
胰腺肿瘤%腺癌%Numb%鼠双微体2%p53
胰腺腫瘤%腺癌%Numb%鼠雙微體2%p53
이선종류%선암%Numb%서쌍미체2%p53
Pancreatic neoplasms%Adenocarcinoma%Numb%MDM2%p53
目的 探讨胰腺导管腺癌(PDAC)中Numb、MDM2和p53表达相关性及临床病理学意义.方法 应用免疫组化法检测65例配对的PDAC及癌旁组织石蜡标本中Numb、MDM2和p53蛋白表达情况,观察三者表达相关性及与患者临床病理学参数的关系;再以Western blot法检测16对配对新鲜PDAC组织及癌旁组织中三者的表达水平.通过免疫荧光法、Western blot法和实时定量(qRT)-PCR法检测Numb在不同分化的胰腺癌细胞株Capan-2、PANC-1及AsPC-1中的表达.分别采用配对t检验、x2检验、Kaplan-Meier、Cox回归分析等统计学方法分析实验结果.结果 免疫组化检测结果显示,Numb在PDAC及配对癌旁组织中表达差异无统计学意义(t=1.746,P=0.086);MDM2和p53在PDAC中表达高于癌旁组织(t=3.294,P=0.002;t=3.152,P=0.002);Numb表达与肿瘤大小(x2=5.206,P=0.023)、分化程度(x2=7.802,P=0.005)及UICC分期(x2=4.770,P=0.029)呈负相关;MDM2和p53表达分别与胰腺癌T及UICC分期呈正相关(x2=5.182,P=0.023;x2=6.448,P=0.011).相关性分析结果显示,Numb与MDM2呈负相关(r=-0.283,P=0.023),但他们与p53表达无明显相关性(P＞0.05).单因素和多因素分析结果显示,Numb是影响胰腺癌患者预后的一个保护因素(x2=5.408,P=0.020),并且Numb阳性和MDM2阴性表达的胰腺癌患者预后更好(x2=5.868,P=0.015).Western blot检测结果显示,Numb在8例高分化PDAC中蛋白表达高于配对的癌旁组织(t=1.092,P=0.020),而MDM2和p53在16例PDAC中呈明显高表达(t=3.263,P=0.005;t=3.607,P=0.003).免疫荧光、Western blot和qRT-PCR检测结果显示,随着胰腺癌细胞分化程度增高,Numb荧光强度、蛋白和mRNA表达水平逐渐增强.结论 Numb在胰腺癌发生发展中主要充当一个抑癌基因的角色,Numb、MDM2和p53可能共同参与胰腺癌的发生进展.
目的 探討胰腺導管腺癌(PDAC)中Numb、MDM2和p53錶達相關性及臨床病理學意義.方法 應用免疫組化法檢測65例配對的PDAC及癌徬組織石蠟標本中Numb、MDM2和p53蛋白錶達情況,觀察三者錶達相關性及與患者臨床病理學參數的關繫;再以Western blot法檢測16對配對新鮮PDAC組織及癌徬組織中三者的錶達水平.通過免疫熒光法、Western blot法和實時定量(qRT)-PCR法檢測Numb在不同分化的胰腺癌細胞株Capan-2、PANC-1及AsPC-1中的錶達.分彆採用配對t檢驗、x2檢驗、Kaplan-Meier、Cox迴歸分析等統計學方法分析實驗結果.結果 免疫組化檢測結果顯示,Numb在PDAC及配對癌徬組織中錶達差異無統計學意義(t=1.746,P=0.086);MDM2和p53在PDAC中錶達高于癌徬組織(t=3.294,P=0.002;t=3.152,P=0.002);Numb錶達與腫瘤大小(x2=5.206,P=0.023)、分化程度(x2=7.802,P=0.005)及UICC分期(x2=4.770,P=0.029)呈負相關;MDM2和p53錶達分彆與胰腺癌T及UICC分期呈正相關(x2=5.182,P=0.023;x2=6.448,P=0.011).相關性分析結果顯示,Numb與MDM2呈負相關(r=-0.283,P=0.023),但他們與p53錶達無明顯相關性(P＞0.05).單因素和多因素分析結果顯示,Numb是影響胰腺癌患者預後的一箇保護因素(x2=5.408,P=0.020),併且Numb暘性和MDM2陰性錶達的胰腺癌患者預後更好(x2=5.868,P=0.015).Western blot檢測結果顯示,Numb在8例高分化PDAC中蛋白錶達高于配對的癌徬組織(t=1.092,P=0.020),而MDM2和p53在16例PDAC中呈明顯高錶達(t=3.263,P=0.005;t=3.607,P=0.003).免疫熒光、Western blot和qRT-PCR檢測結果顯示,隨著胰腺癌細胞分化程度增高,Numb熒光彊度、蛋白和mRNA錶達水平逐漸增彊.結論 Numb在胰腺癌髮生髮展中主要充噹一箇抑癌基因的角色,Numb、MDM2和p53可能共同參與胰腺癌的髮生進展.
목적 탐토이선도관선암(PDAC)중Numb、MDM2화p53표체상관성급림상병이학의의.방법 응용면역조화법검측65례배대적PDAC급암방조직석사표본중Numb、MDM2화p53단백표체정황,관찰삼자표체상관성급여환자림상병이학삼수적관계;재이Western blot법검측16대배대신선PDAC조직급암방조직중삼자적표체수평.통과면역형광법、Western blot법화실시정량(qRT)-PCR법검측Numb재불동분화적이선암세포주Capan-2、PANC-1급AsPC-1중적표체.분별채용배대t검험、x2검험、Kaplan-Meier、Cox회귀분석등통계학방법분석실험결과.결과 면역조화검측결과현시,Numb재PDAC급배대암방조직중표체차이무통계학의의(t=1.746,P=0.086);MDM2화p53재PDAC중표체고우암방조직(t=3.294,P=0.002;t=3.152,P=0.002);Numb표체여종류대소(x2=5.206,P=0.023)、분화정도(x2=7.802,P=0.005)급UICC분기(x2=4.770,P=0.029)정부상관;MDM2화p53표체분별여이선암T급UICC분기정정상관(x2=5.182,P=0.023;x2=6.448,P=0.011).상관성분석결과현시,Numb여MDM2정부상관(r=-0.283,P=0.023),단타문여p53표체무명현상관성(P＞0.05).단인소화다인소분석결과현시,Numb시영향이선암환자예후적일개보호인소(x2=5.408,P=0.020),병차Numb양성화MDM2음성표체적이선암환자예후경호(x2=5.868,P=0.015).Western blot검측결과현시,Numb재8례고분화PDAC중단백표체고우배대적암방조직(t=1.092,P=0.020),이MDM2화p53재16례PDAC중정명현고표체(t=3.263,P=0.005;t=3.607,P=0.003).면역형광、Western blot화qRT-PCR검측결과현시,수착이선암세포분화정도증고,Numb형광강도、단백화mRNA표체수평축점증강.결론 Numb재이선암발생발전중주요충당일개억암기인적각색,Numb、MDM2화p53가능공동삼여이선암적발생진전.
Objective To study the relationship and clinicopathological significance of Numb,MDM2 and p53 expression in human pancreatic cancer.Methods The expression of Numb,MDM2 and p53 proteins in 65 cases of paired paraffin embedded pancreatic ductal adenocarcinoma (PDAC) specimens and adjacent non-cancerous pancreas was detected by immunohistochemistry (IHC).The relationship among their expression and clinicopathological characters was analyzed.Westem blot was used to examine their expression in 16 paired fresh PDAC specimens and adjacent non-cancerous pancreatic tissues.Meanwhile,Numb expression in Capan-2,PANC-1 and AsPC-1 pancreatic cancer cells with different differentiation were detected by immunofluorescence (IF),Westem blot and quantitative real-time (qRT)-PCR,respectively.Paired sample t-test,x2 test,Kaplan-Meier and Cox regression were used to analyze the results of our experiments,respectively.Results IHC showed that there was no differential expression of Numb in PDAC and adjacent pancreas (t =1.746,P =0.086),while the expression of MDM2 and p53 was significantly increased in PDAC,compared to that in paired normal pancreas (t =3.294,P =0.002 ; t =3.152,P =0.002,respectively).Numb expression was negatively associated with tumor size (x2 =5.206,P =0.023),differentiation (x2 =7.802,P =0.005) and UICC stage (x2 =4.770,P =0.029),while expression of MDM2 and p53 was positively associated with tumor T and TNM stage,respectively (x2 =5.182,P =0.023 ; x2 =6.448,P =0.011).Correlation analysis showed a negative association between Numb and MDM2(r =-0.283,P =0.023),but there was no relationship of them with p53(P ＞0.05).Univariate and multivariate analysis revealed that Numb was a protective prognostic indicator for patients with PDAC (x2 =5.408,P =0.020).Moreover,patients with Numb positive and MDM2 negative expression had a significantly better overall survival(x2 =5.868,P =0.015).Western blot showed that Numb expression was much higher in well differentiated PDAC than that in paired normal pancreas (t =1.092,P =0.020),while the expression of MDM2 and p53 was significantly increased in 16 cases of PDAC(t =3.263,P =0.005 ;t =3.607,P =0.003,respectively).Numb expression was gradually increased in pancreatic cancer cells with the increasing degree of cell differentiation detected by IF,Westem blot and qRT-PCR.Conclusions Numb acts as a tumor suppressor gene in the development of PDAC.Numb,MDM2 and p53 might coordinately participate in the development of PDAC.
