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kai1 inhibits lymphangiogenesis and lymphatic metastasis of pancreatic cancer in vivo

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Hepatobiliary & Pancreatic Diseases International 2014年第1期13卷 87-92页

作者： Xu Liu Xiao-Zhong Guo Hong-Yu Li Jiang Chen Li-Nan Ren Chun-Yan Wu Department of Gastroenterology General Hospital of Shenyang Military Area

BACKGROUND: Several studies have shown that kai1 inhibits tumor metastasis, but its mechanism is not clear. The present study aimed to determine the role of kai1 in lymphatic metastasis, specifically in pancreatic cancer. METHODS: The kai1 gene was transfected into the pancreatic cancer cell line MIA PaCa-2 and PANC-1 by using liposomes and selected by G418, and the protein was measured by Western blotting. After successful infection, the cell growth curve was studied by MTT, vascular endothelial growth factor C(VEGF-C) secretion by pancreatic cancer cell were measured by ELISA. The kai1 and pCMV transfected MIA PaCa-2 cells were renamed as MIA PaCa-2-K and MIA PaCa-2-p. These two kinds of cells were injected into the subcuticular layer of nude mice; both tumor growth and metastasis through the lymphatic nodes were assessed. Lymphangiogenesis in tumors was measured by immunohistochemistry. RESULTS: The VEGF-C secretion was significantly reduced in MIA PaCa-2 cells compared with PANC-1 cells after being transfected with the kai1 gene. The growth rate of subcutaneous tumors was similar after the injection of MIA PaCa-2-K, MIA PaCa-2, and MIA PaCa-2-p. MIA PaCa-2-K tumors showed slower lymphangiogenesis and lymph node metastasis compared with MIA PaCa-2 and MIA PaCa-2-p tumors. CONCLUSION: The overexpression of kai1 inhibits the lymphangiogenesis and lymph node metastasis of MIA PaCa-2 pancreatic tumors.

关键词： kai1 pancreatic cancer lymphatic metastasis lymphangiogenesis

Expression of nm23, kai1 and spiral computed tomography findings in primary gallbladder carcinoma

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Chinese Medical Journal 2009年第21期122卷 2666-2668页

作者： JIANG Wen-xia SONG Bo-gen WANG Pei-jun Department of Pathology Medical College of Tongji University Shanghai 200092 China Department of Radiology Affiliated Tongji Hospital of Tongji University Shanghai 200065 China

Primary gallbladder carcinoma is the most common malignant tumor of the biliary system. There is a reduced likelihood for radical cure of primary gallbladder carcinomas because of the insidious onset.1 Spiral computed tomography （SCT） scanning is an important means for diagnosing lesions involving the gallbladder. We determined the expression of anti-metastasis protein nm23 and kai1 using immunohistochemical method and analyzed the relationship between their expression and the pre-operatively SCT imaging findings, the histopathologic grade, clinical stage, 5-year survival rate in 35 cases of primary gallbladder cancer.

关键词： gallbladder carcinoma spiral computed tomography nm23 kai1

Expression and clinical significance of p53,JunB and kai1/CD82 in human hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2009年第4期8卷 389-396页

作者： Guo, Cheng Liu, Qing-Guang Zhang, Lei Song, Tao Yang, Xue Xi An Jiao Tong Univ Sch Med Affiliated Hosp 1 Dept Hepatobiliary Surg Xian 710061 Peoples R China

BACKGROUND: The mechanism of regulation of kai1, a specific tumor metastasis suppression gene, is controversial. A recent study showed that the synergism of wild-type p53 and JunB has the function of regulating the expression of kai1, a metastasis inhibiting factor in prostate cancer cells. The wild-type p53 gene is an activator of apoptosis and is closely related to malignant tumor cell multiplication. JunB, a member of the fos/jun family, is a key component of activator protein transcription factor and a major target element in the transmission pathway of mitosis. This study aimed to evaluate the relationship between the expression of kai1 and p53 combined with JunB in tumor tissues and clinical outcomes in hepatocellular carcinoma (HCC) patients. METHODS: Quantitative real-time RT-PCR, Western blotting techniques and immunohistochemistry were used to evaluate the expression of kai1 mRNA, kai1/CD82, p53 and JunB in HCC patients, and the relationship between their expression and the clinicopathological prognostic parameters was analyzed. RESULTS: In cancer tissues, the values for positive expression of kai1 mRNA, kai1/CD82, p53 and JunB were 31.25%, 26.25%, 48.75%, and 20.00%, respectively, while in adjacent non-tumor tissues, they were 100%, 94.74%, 2.63%, and 76.32%, respectively. There was no correlation between the expression levels of p53 or JunB and kai1 mRNA or kai1/CD82. However, there were significant correlations between the expression levels of p53 combined with JunB and not only kai1 mRNA but also kai1/CD82 proteins. CONCLUSIONS: When p53 dysfunction and low expression of JunB are simultaneous, they may play an important role in down-regulating the expression of kai1 by synergism in HCC. But further studies in vivo and in vitro are needed to verify these results.

关键词： hepatocellular carcinoma JunB kai1 p53 gene expression prognosis

Double Lethal Effects of Fusion Gene of Wild-type p53 and JunB on Hepatocellular Carcinoma Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第5期32卷 663-668页

作者：郭成刘青光张雷杨雪宋涛姚英民 Department of Hepatobiliary Surgery the First Affiliated HospitalSchool of MedicineXi'an Jiaotong University

This study explored the double lethal effects of pEGFP-C1-wtp53/junB fusion gene on hepatocellular carcinoma (HCC) cells.wtp53/junB fusion gene was constructed and transformed into HepG2 cell line.Expression of kai1 was detected by quantitative real-time PCR and Western blotting,cells apoptosis rate was detected by flow cytometry,proliferation of cells was detected by MTT chromometry,cell transmigration was detected by using transwell systems.The results showed that after transformation with pEGFP-C1-wtp53/JunB,the expression level of kai1 protein was up-regulated,being 8.13 times the blank control group in HepG2 cells and significantly higher than 2.87 times which transformed with pEGFP-C1-JunB,3.11 times which transformed with pEGFP-C1-wtp53 (P1).Apoptosis rate of HepG2 cells transformed with pEGFP-C1-wtp53/JunB was significantly higher than that of other groups (P1),and invasive ability of HepG2 cells transformed with pEGFP-C1-wtp53/JunB was significantly lower than other groups (P1).It was concluded that the fusion gene of wtp53 and JunB could not only inhibit the growth of hepatoma cells and promote tumor cell apoptosis,but also suppress the invasive ability of tumor cells by up-regulating the expression of kai1.

关键词： hepatocellular carcinoma kai1 CD82 p53 JunB

kai1 gene is differently expressed in papillary and pancreatic cancer:influence on metastasis

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World Journal of Gastroenterology 2000年第6期6卷 866-871页

作者： Helmut Friess Markus W.Buchler Department of Visceral and Transplantation Surgery University of BernInselspitalBernSwitzerland

AIM To compare kai1 in cancer of papilla ofVater and pancreas to evaluate whether there aredifferences in biologic behavior which mightaccount for prognosis.METHODS We compared the expression in 24papillay and 29 pancreatic cancers usingNorthern blot analysis,immunochemical assayand in situ hybridization,and investigatedwhether early diagnosis or molecular differencespredict the outcome in these tumor entities.RESULTS By Northern blot analysis there is nostatistical difference of kai1 levels in normaland cancerous papilla.No association betweenkai1 mRNA expression and tumor stage or tumordifferentiation was found in the tumors.Byimmunohistochemical assay,kai1 staining incytoplasm of papillary cancer cells was similarto that of normal papillary cells.By in situhybridization,the results of kai1 mRNAexpression in normal and cancerous papilla weresimilar to those with immunohistochemicalassay.The normal and cancerous pancreastissues were also analyzed by the methods usedin papillary samples.CONCLUSION Although the biologic roles of kai1 have not been clarified, our results suggest that kai1 may restrict the progression of malignant papillary cancer, but its expression might not have any effect on the characteristics of papillary tumor, whereas by the analysis of KAl1 gene, its reduced expression is closely related to the progression and metastases of pancreatic cancer.

关键词： pancreatic neoplasms papillary neoplasms kai1 gene immunohistochemistry in situ hybridization blotting northern

kai1 is a potential target for anti-metastasis in pancreatic cancer cells

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World Journal of Gastroenterology 2008年第7期14卷 1126-1132页

作者： Jian-Hua Xu Xiao-Zhong Guo Li-Nan Ren Li-Chun Shao Min-Pei Liu Department of Gastroenterology The General Hospital of Shenyang Military Command Shenyang 110016 Liaoning Province China

AIM: To investigate whether kai1, as a metastasis suppressor gene, is associated with invasive and metastatic ability of pancreatic cancer cells.METHODS: kai1 gene was transfected into pancreatic cancer cell line MiaPaCa Ⅱ by liposomes selected with G418. Expression of transfected cells was measured by Western blotting, immunofluorescence and immunocytochemistry. Tumor cell invasion and metastatic ability were detected through gelatinase activity and reconstituted basement membrane (Matrigel) assay. pCMV-kai1 was directly injected into the heterotopic human pancreatic adenocarcinoma successfully established in the groin of BALB/C nude mice, by subcutaneous injection of MiaPaCa Ⅱ pancreatic cancer cells. The statistical analysis between groups was determined by Student's two tailed t test.RESULTS: By Western blotting, MiaPaCa Ⅱ cells transfected by kai1 gene indicated kai1 expression at approximately 29.1 kDa. Cytoplasm staining was positive and uniformly spread in transfected cancer cells, using immunohistochemistry and immunofluorescence. The most obvious difference was present after 30 h (MiaPaca Ⅱ 43.6 ± 9.42, pCMV-MiaPaca Ⅱ 44.8 ± 8.56, pCMV-kai1-MiaPaca Ⅱ 22.0 ± 4.69, P 1 ± 0.65, P kai1 transfectants with kai1-Lipofectamine 1.22 ± 0.31 in A group were lower than control 4.61 ± 1.98 and pCMV-kai 11.67 ± 0.81. Analyses of metastases with and without kai1 transfection in mice were different in liver and lung between controls 1.62 ± 0.39, 0.45 ± 0.09, pCMV-kai 1.01 ± 0.27, 0.33 ± 0.09 and kai1-Lipofectamine 0.99 ± 0.21, 0.30 ± 0.09 respectively (P kai1 gene was found in transfected MiaPaCa Ⅱ human pancreatic cancer cells with lower metastatic ability. kai1 gene plays an important role in inhibiting metastasis of pancreatic cancer after direct injection into pa

关键词： kai1 胰腺癌基因转染肿瘤细胞

kai 1通过Integrin β1调控乳腺癌细胞生物学行为的研究

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KAI 1通过Integrin β1调控乳腺癌细胞生物学行为的研究

作者：孙檬大连医科大学

学位级别：硕士

研究背景乳腺癌(Breast cancer,BC)目前已成为全球第一大新增癌症病例,据2020年世界卫生组织国际癌症研究机构(IARC)最新数据统计,2020年全球新增癌症病例约1930万,其中女性乳腺癌占11.7%,在数量上已经首次超越了肺癌(11.4%),成为全球... 详细信息

研究背景乳腺癌(Breast cancer,BC)目前已成为全球第一大新增癌症病例,据2020年世界卫生组织国际癌症研究机构(IARC)最新数据统计,2020年全球新增癌症病例约1930万,其中女性乳腺癌占11.7%,在数量上已经首次超越了肺癌(11.4%),成为全球发病率最高的癌症。在过去的30年中,我国女性乳腺癌的发病人数和死亡人数持续递增,已成为威胁女性健康与生命的头号敌人。kai1(Kang Ai 1/CD82)属于跨膜4超家族(TM4SF)成员,是在男性前列腺癌细胞11号染色体上分离、克隆出来的一种新的肿瘤转移抑制因子,其表达下调或缺失与多种恶性肿瘤的进程相关。整合素β1(Integrinβ1)是整合素家族成员之一,介导细胞与细胞、细胞与基质间的反应,癌症干细胞中整合素β1的表达升高。近年来有研究结果显示kai1与Integrinβ1在肺癌中的表达呈负相关。在乳腺癌细胞中,kai1与Integrinβ1二者的关系及其作用尚不十分清楚。目的1.检测乳腺癌组织及正常乳腺组织中kai1及Integrinβ1蛋白的表达情况,探讨二者的表达是否存在相关性。2.检测kai1敲低前后乳腺癌细胞中Integrinβ1蛋白的表达情况,探讨下调kai1对乳腺癌细胞侵袭、迁移等生物学行为的影响。3.探讨kai1能否与Integrinβ1结合以及kai1能否对Integrinβ1的表达进行调控。方法1.对48例乳腺癌标本采用免疫组织化学染色法,检测乳腺癌组织与周围正常乳腺组织中kai1和Integrinβ1的表达情况,用统计学方法计算表达的阳性率。2.采用非参数Spearman相关性分析在正常乳腺组织和乳腺癌组织中,kai1与Integrinβ1表达的相关性。3.选用人乳腺癌细胞株(MDA-MB-231、MCF-7),实验分成si-kai1基因转染组和对照组(NC),下调kai1后,采用Western blot实验分别检测两组乳腺癌细胞中kai1、Integrinβ1蛋白的表达情况,检测MMP2、MMP9的表达变化;运用Transwell小室实验,使用Image J软件统计细胞穿膜数量;采用划痕实验,观察乳腺癌细胞在24h、48h的迁移情况,使用Image J软件统计划痕宽度。4.用对照Ig G抗体、kai1抗体或Integrinβ1抗体免疫沉淀MCF-7细胞裂解物,用Western blot实验分别检测co-IP组和Ig G组中kai1和Integrinβ1的蛋白印迹。结果1.对48例乳腺癌标本采用免疫组织化学染色法,检测发现,与正常乳腺组织比较,乳腺癌组织中kai1表达低于正常组织,Integrinβ1表达高于正常组织。2.采用非参数Spearman相关性分析在正常乳腺组织和乳腺癌组织中,kai1与Integrinβ1的表达呈负相关。3.在人乳腺癌细胞株(MDA-MB-231、MCF-7)中,瞬时转染下调kai1后,Western blot实验结果显示,实验组kai1的表达水平较对照组降低,Integrinβ1的表达水平较对照组升高;实验组MMP2、MMP9的表达水平高于对照组;Transwell小室结果显示,实验组细胞穿膜数量高于对照组;划痕实验结果显示,实验组划痕愈合能力高于对照组。4.用对照Ig G抗体、kai1抗体或Integrinβ1抗体免疫沉淀MCF-7细胞裂解物,Western blot实验检测kai1和Integrinβ1的蛋白印迹,co-IP组均检测到kai1和Integrinβ1条带,Ig G组未检测到蛋白印迹。结论1.kai1在乳腺癌组织中表达下降,Integrinβ1在乳腺癌组织中表达升高,两者呈负相关。2.下调kai1促进乳腺癌细胞的侵袭和迁移。3.kai1能够与Integrinβ1结合。4.kai1能够调控Integrinβ1的表达。5.提示kai1能通过负向调控Integrinβ1的表达从而抑制乳腺癌细胞的侵袭和转移,kai1有望作为乳腺癌治疗和预后评估的新靶标。

关键词：乳腺癌 kai1 Integrinβ1 mmp2 mmp9 乳腺癌细胞系(MCF-7和MDA-MB-231)

胃腺癌组织kai1的表达及与血管、淋巴管生成的关系

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四川大学学报（医学版） 2014年第1期45卷 43-48页

作者：周蕾武世伍俞岚宋文庆承泽农王丹娜蚌埠医学院第一附属医院病理科蚌埠医学院病理学教研室蚌埠233003

目的寻找能预测胃腺癌(gastric adenocarcinoma,GAC)浸润、转移及预后的指标。方法采用免疫组织化学法检测145例胃腺癌和50例癌旁正常胃黏膜组织抗癌1号(kai1)、CD34及D2-40的表达。通过CD34的阳性表达计数微血管密度(microvessel densi... 详细信息

目的寻找能预测胃腺癌(gastric adenocarcinoma,GAC)浸润、转移及预后的指标。方法采用免疫组织化学法检测145例胃腺癌和50例癌旁正常胃黏膜组织抗癌1号(kai1)、CD34及D2-40的表达。通过CD34的阳性表达计数微血管密度(microvessel density,MVD),通过D2-40的阳性表达计数淋巴管密度(lymph vessel density,LVD)。分析kai1、MVD及LVD之间的相互关系,以及它们与胃腺癌浸润、转移及预后的关系。结果在癌旁正常胃黏膜组织中,kai1的阳性表达率、MVD和LVD分别为92.0%、(9.2±7.8)/低倍镜(LP)和(7.5±7.6)/LP;在GAC组织中,kai1、MVD和LVD分别为37.2%、(21.6±9.1)/LP和(22.6±12.7)/LP,两组之间差异均有统计学意义(Pkai1的表达水平、MVD及LVD的高低与肿瘤细胞的pTNM分期、浸润深度、淋巴结转移与否有关(Pkai1的表达与MVD及LVD呈负相关,MVD与LVD呈正相关(P均kai1的表达、MVD及pTNM分期是影响GAC患者术后生存时间的独立因素。Kaplan-Meier生存分析表明,MVD≥22组和LVD≥23组患者其术后5年生存率低于MVD1);kai1表达阳性的患者其术后5年生存率高于阴性患者(P1)。结论早期联合检测kai1、CD34及D2-40对GAC的浸润、转移及预后判断有重要意义。

关键词：胃腺癌 kai1 微血管密度淋巴管密度预后

液相色谱-质谱法对鼠胰腺癌组织中氧化脂质代谢分析及其与转移相关机制的研究

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液相色谱-质谱法对鼠胰腺癌组织中氧化脂质代谢分析及其与转移相...

作者：姚莉颖锦州医科大学

学位级别：硕士

目的胰腺癌(Pancreatic Cancer,PC)是一种病死率极高的恶性肿瘤,其高死亡率、低生存率及预后极差的突出特征主要归结于PC的高转移性。早期诊断是治疗乃至治愈PC的关键,而探索与PC相关的新生标志物的研究能够给PC的早期诊断以及靶向治疗... 详细信息

目的胰腺癌(Pancreatic Cancer,PC)是一种病死率极高的恶性肿瘤,其高死亡率、低生存率及预后极差的突出特征主要归结于PC的高转移性。早期诊断是治疗乃至治愈PC的关键,而探索与PC相关的新生标志物的研究能够给PC的早期诊断以及靶向治疗带来新的希望。氧化脂质组学是在代谢组学基础上出现的一种新的多组学研究方法,对PC及与其转移相关的氧化脂质进行深入研究,有利于PC诊断模型的建立和PC发生发展及转移机制的探索。目前国内外对此研究领域仍少有相关报道。本研究的主要目的在于通过构建人源性鼠PC动物模型,并基于液相色谱-质谱(Liquid Chromatography-Mass Spectrometry,LC-MS/MS)联合代谢组学平台检测,经多元统计分析法明晰各组样本间氧化脂质代谢物谱差异,阐明动物体内条件下PC组织氧化脂质代谢的特点,以及抑癌基因kai1对PC氧化脂质代谢的可能影响,进而探讨氧化脂质体作为PC早期诊断生物标志物的可行性。方法构建人源性鼠PC动物模型,选取9只4-6周龄雌性BALB/c裸鼠随机分3组,每组分别在胰腺原位注射PANC-1细胞悬液、kai1-PANC-1细胞悬液、脾脏注射PANC-1-ATX shRNA细胞悬液,待成瘤后解剖取材。应用液相色谱-质谱联用法检测各组PC组织及转移瘤组织中小分子氧化脂质代谢物的表达变化情况。对检测获得数据行主成分分析(Principal Components Analysis,PCA)和正交偏最小二乘法判别分析(Orthogonal Partial Least Squares Discriminant Analysis,OPLS-DA),联合京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库筛选并鉴定出差异代谢物。利用分层聚类分析以及代谢通路富集,研究其可能参与的代谢通路。结果1、基于LC-MS/MS代谢组学平台,以及PCA、OPLS-DA等多元统计学方法对氧化脂质代谢物进行轮廓分析,发现建立的各组模型质量良好,相关数据整体呈现组间离散,组内聚集的态势;2、在全部的9个差异配对组中(ATX-PC vs ATX-LC、kai1-LC vs ATX-LC、kai1-PC vs ATX-PC、kai1-PC vs kai1-LC、NC-LC vs ATX-LC、NC-LC vs kai1-LC、NC-PC vs ATX-PC、NC-PC vs kai1-PC和NC-PC vs NC-LC),共筛查出64种呈现差异表达的氧化脂质代谢物,并且全部64种氧化脂质代谢物本身皆属于不饱和脂肪酸成分;3、通过对kai1-LC vs ATX-LC与NC-LC vs ATX-LC两个差别配对组的氧化脂质代谢谱进行比较,发现前列腺素类分子(PGE2、PGD2)在敲除ATX的肝转移组织中的含量均呈现明显下降的趋势,以及在PC肝转移组织中,ATX敲除可能会通过引起下游LPA的下调导致前列腺素类分子水平下调从而参与肿瘤的转移;4、通过KEGG通路分析并结合Student-t检验,发现NC-PC vs kai1-PC组中的亚麻酸(Linolenic Acid,LA)代谢具有统计学意义(p≤0.05),其中20个氧化脂质代谢物存在显著差异(19个上调,1个下调),差异代谢物分别属于花生四烯酸(Arachidnic Acid,AA)、LA、二十碳五烯酸(Eicosapentaenoic Acid,EPA)和二十二碳六烯酸(Docosahexaenoic Acid,DHA)成分。生信分析显示上调表达的氧化脂质代谢物12(13)-EpOME参与了kai1/CD82抑制PC转移过程。结论氧化脂质代谢参与PC的发生、发展及转移进程。前列腺素类分子(PGE2、PGD2)通过ATX-LPA轴促进PC肝转移的进程;氧化脂质代谢物12(13)-EpOME参与了kai1/CD82基因对PC转移的抑制过程,该氧化脂质代谢物可能成为PC早期诊断的潜在生物标志物,有必要对其进行深入的研究。

关键词：胰腺癌氧化脂质代谢组学 kai1 人源性小鼠胰腺癌模型