Study On The Expression Of Marker Proteins Of Cancer Stem Cell And The Metastatic And Invasive Mechamism In The Benign And Malignant Lesions Of Gullbladder

Part One Cancer Stem Cells and Their MarkersObjective To study the expression levels of prostate stem cell antigen(PSCA),Oct-4,CD24 and CD44V6 in the gallbladder adenocarcinoma,perirumoral tissues,adenomatous polyp,and chronic cholecystitis,and to detect their clinicopathologic significances in gallbladder adenocarcinoma.Methods Immunohistocemical method of EnVision^TM(two steps)for PSCA,Oct-4,CD24 and CD44V6 was used in the routinely paraffin-embedded sections(after 4%formaldehyde fixing)of specimens from gallbladder adenocarcinoma(n=108),peritumoral tissues(n=46), adenomatous polyp(n=15),and chronic cholecystitis(n=35)Results(1) In the 108 cases of gallbladder adenocarcinoma,PSCA, Oct-4,CD24 and CD44V6 positive cases were respectively 59(54.6%),60 (55.6%),60(55.6),57(52.8%),and 60(55.6%),and their scores were respectively2.09±1.94,2.06±1.86,2.43±1.65,and 2.52±1.60.In the 46 cases of perirumoral tissues,PSCA,Oct-4,CD24 and CD44V6 positive cases were respectively 9(19.6%);10(22.5%);8(17.4%),and 9 (19.6%),their scores were respectively 0.82±1.11 0.85±1.02 0.78±1.36 and 0.82±1.39.In the 15 cases of adenomatous polyp,PSCA,Oct-4,CD24 and CD44V6 positive cases were respectively 4(26.7%),3(20%),and 4 (26.7%) and their scores were respectively 0.74±1.25,0.69±1.32, 0.72±1.35,and 0.78±1.28 In the 35 cases of chronic cholecystitis,PSCA,oct-4,CD24,and.CD44V6 positive cases were respectively 5(14.3%) 5(14.3%) 4(14.4%) and 6(17.1%),and their scores were respectively 0.68±1.21,0.67±0.95,0.54±1.44,and 0.58±1.46, The gallbladder epithelium of PSCA,Oct-4,CD24 and CD44V6 positive cases all showed middle to severely-atypical hyperplasia.The positive rates of PSCA,ct-4,CD24 and CD44V6 expressions and their scores in gallbladder adenocarcinoma were significantly higher than those in peritumoral tissues(PSCA,x²=10.09,P＜0.01,Ï„=4.23,P＜0.01;Oct-4, x²=14.88,P＜0.01,Ï„=4.32,P＜0.01;CD24,x²=16.58,p＜0.01,t=5.89, P＜0.01;CD44V6,x²=16.90,P＜0.01,Ï„=6.07,P＜0.01),adenomatous polyp (PSCA,x²=4.12,P＜0.05;Ï„=2.60P＜0.05 Oct-4,x²=6.66,p＜0.01,Ï„=2.80, P＜0.01;CD24,x²=5.66,P＜0.05,Ï„=3.78,P＜0.01;CD44V6,x²=4.12, P＜0.05,Ï„=4.05,P＜0.01),and chronic cholecystitis(PSCA,x²=17.40, P＜0.01,Ï„=6.10,P＜0.01;Oct-4,x²=18.16,p＜0.01,Ï„=4.21,P＜0.01;CD24, x²=18.48,P＜0.01,Ï„=6.10,P＜0.01;CD44v6,x²=15.69,P＜0.01,Ï„=6.62, P＜0.01).(2)The positive rates of PSCA,Oct-4,CD24 and CD44V6 expressions and their scores were significantly lower in the cases of adenomatous canceration or well-differentiated adenocarcinoma,maximal diameter of mass＜2cm,no-metastasis of lymph node,and no-invasion of regional tissues than those in the ones of pooly-differentiated adenocarcinoma, maximal diameter of mass≥2cm,metastasis of lymph node and invasion of regional tissues(P＜0.05,or P＜0.01).(3)Highly consistencies in PSCA,Oct-4,CD24 and CD44V6 expressions were found in gallbladder adenocarcinoma(P＜0.01),and significantly positive correlations were found among their scores(PSCA vs Oct-4,r=0.44,P＜0.01;PSCA vs CD24,r=0.36,P＜0.01;PSCA vs CD44V6,r=0.49,P＜0.01;Oct-4 vs CD24,r=0.51,P＜0.01;Oct-4 vs CD44V6,r=0.39,P＜0.01;CD24 vs CD44v6,r=0.48,P＜0.01).Conclusions PSCA,Oct-4,CD24 and CD44v6,as markers of cancer stem cells,played an important role in the carcinogenesis, progression,metastatic potency,invasive capability of gallbladder adenocarcinoma,and positive or high expression ones had poor prognostis The cancer stem cells in the cancer tissues might play an important role in the genesis,progression,biological behavior and prognosis of gallbladder adenocarcinoma.Detecting the PSCA,Oct-4,CD24 and CD44V6 levels in the benign lesions of gallbladder might have significant clinical values for the prevention and early-finding of gallbladder adenocarcinoma. Part Two the Controlling Mechanism of Metastasis and Invasion of Gallbladder AdenocarcinomaChapter One Microvessels counts and lymph vessels counts and their significances in benign and malignant lesions of gallbladderObjective To study the microvessel(MV)counts,and lymph vessel (LV)counts in gallbladder adenocarcinoma,peritumoral tissues, adenomatous polyp,and chronic cholecystitis,and to detect their clinicopathologic significances in gallbladder adenocarcinoma.Methods Streptavidin-peroxidase immunohistochemical methods for MV and LV was used in the routinely paraffin-embedded sections(after 4%formaldehyde fixing)of specimens from gallbladder adenocarcinoma(n=108),peritumoral tissues(n=46),adenomatous polyp(n=15),and chronic cholecystitis(n=35).Selected ten regions tha MV and LV were most dense under the low-power field,and then counted the numbers of MV and LV of the ten regions under the high-power microscope;the average of the numbers was the counts of MV and LV of the case.Results The counts of MV and LV(64.2±11.6,11.4±5.2)in gallbladder adenocarcinoma were significantly higher than those in peritumoral tissues(28.2±12.4,Ï„=17.6,P＜0.01;6.8±6.2,Ï„=4.63,P＜0.01), adenomat-ous polyp(32.4±11.8,Ï„=9.94,P＜0.01;5.6±2.9,Ï„=4.17, P＜0.01),and chronic cholecystitis(22.2±11.3,Ï„=18.26,P＜0.01;5.1±3.9, Ï„=6.63,P＜0.01).The counts of MV and LV were significantly lower in the cases of adenomatous canceration or well-differentiated adenocarcinoma,maximal diameter of mass＜2cm,no-metastasis of lymph node,and no-invasion of regional tissues than those in the ones of poorly-differentiated adenocarcinoma,maximal diameter of mass≥2cm, metastasis of lymph node,and invasion of regioonal tissues(P＜0.01). Significantly positive correlation was found between MV counts and LV counts in gallbladder adenocarcinoma(r=0.38,p＜0.01).Conclusions MV counts and LV counts might be important biological markers reflecting the progression,clinical behavior and prognosis of gallbladder adenocarcinoma.Chapter Two Study of metastatic controlling genesObjective To study the expressions of KAI-1mRNA, Kiss-1mRNA,MTA1 and CD146 in gallbladder adenocarcinoma,peritumoral tissues,adenomatous polyp,and chronic cholecystitis,and to detect their clinico- pathologic significances in gallbladder adenocarcinoma.Methods In situ hybridization for staining KAI-1mRNA and Kiss-1mRNA,and immunohistochemical method of EnVision^TM(two steps)for MTA1 and CD146 were used in the routinely paraffin-embedded sections(after 4%formaldehyde fixing) of specimens from gallbladder adenocarcinoma(n=108),peritumoral tissues(n=46),adenomatous polyp(n=15),and chronic cholecystitis(n=35).Results The positive rates of KAI-1mRNA and Kiss-1mRNA (51.9%,53.7%)were significantly lower in gallbladder adenocarcinoma than those in peritumoral tissues(KAI-1mRNA,80.4%,x²=11.02, P＜0.01;Kiss-1mRNA,82.6%,x²=11.48,P＜0.01),adenomatous polyp(KAI-1mRNA,86.7%,x²=6.48,P＜0.05;Kiss-1mRNA,86.7%,x 2=5.86,P＜0.05);and chronic cholecystitis(KAI-1mRNA 85.7%,x²=12.64, P＜0.01;Kiss-1mRNA,85.7%,x²=11.44,P＜0.01).The positive rates of MTA1 and CD146(48.1%,53.7%)were significantly higher in gallbladder adenocarcinoma than those in peritumoral tissues(MTA1,26.1%,x 2=6.46,P＜0.05;CD146,30.4%,x²=7.02,P＜0.01),ademomatous polyp(MTA,20.0%,x²=4.22,P＜0.05;CD146,20.0%,x²=8.59%, P＜0.01);and chronic cholecystitis(MTA1,8.6%,x²=13.31,P＜0.01; CD146,5.7%,x²=25.00,P＜0.01).The positive rates of KAI-1mRNA and kiss-1mRNA were significantly higher in cases with adenomafous canceration or well differentiated adenocarcinoma,maximal diameter of regional tissues＜2cm,no-metastasis of lgmph node and no-invasion of regional tissues than in those with poorly-differentiated adenocarcinoma or mucinous adenocarcinoma,maximal diameter of mass≥2cm, metastasis of lymph node and invasion of regional tissues(P＜0.05 or P＜0.01),while MTA and CD146 were on the contrary(P＜0.05 or P＜0.01).The counts of MV and LV in gallbladda adenocarcinoma were significantly lower in KAI-1mRNA and/on kiss-1mRNA positive cases than those in negative cases(P＜0.01),while the counts of MV and LV in gallbladder adenocarcinoma were significantly higher in MTA1,and/or CD146 positive cases than those in negative cases(P＜0.01).Conclusions KAI-1mRNA,kiss-1mRNA,MTA1 and CD146 might be important biological markers reflecting the genesis,progression,clinic biological behavior and prognosis of gallbladder adenocarcinoma,and important markers controlling the metastasis and invasion abilities of gallbladder adenocarcinoma.Their controlling impact might correlate with their controlliny of the angiogenesis and lymphangiolgenesis of tumors.