Gankyrin蛋白在大鼠肝细胞癌变过程中的异常表达及意义

目的探讨肝细胞癌变过程中Gankyrin蛋白的异常表达及其意义。方法127只雄性Wistar大鼠分为对照组(20只)和实验组(107只),实验组大鼠用0.2%二乙基亚硝胺(diethylnitrosamine,DEN)诱导建立肝癌动物模型,应用免疫组化法检测大鼠肝脏在癌变及转移过程中Gankyrin蛋白的表达。结果对照组中未见Gankyrin蛋白表达;实验组中,21例肝硬化中有3例Gankyrin蛋白过表达,68例未转移肝癌中有46例Gankyrin蛋白过表达,18例伴转移肝癌中有17例Gankyrin蛋白过表达。结论Gankyrin蛋白的阳性表达率随病变程度的加重而逐渐升高,这提示Gankyrin蛋白在肝细胞癌形成过程中可能发挥重要的促癌作用,并可能促进了肝癌的转移和高侵袭性发展,其有可能成为肝细胞癌转移早期监测的参考指标。

GST-Gankyrin融合蛋白的原核表达及抗体制备

目的:为进一步研究在肝癌上过表达的癌相关基因Gankyrin的功能,进行GSTGankyrin融合蛋白表达载体的构建、原核表达、及抗体制备.方法:采用逆转录PCR(RTPCR)法从人肝癌细胞系hepG2中扩增GankyrincDNA编码区,并将其重组于谷胱甘肽硫转移酶(GST)融合蛋白表达质粒pGEX4T2中,经酶切、序列鉴定分析后,用该重组质粒转化大肠杆菌DH5α,经IPTG诱导获得表达,并将Gankyrin蛋白免疫家兔,经过Westernblot检测抗Gankyrin抗体的产生.结果:成功构建了Gankyrin原核表达载体,并在大肠杆菌DH5α中获得表达,Westernblot检测证实获得了抗Gankyrin抗体.结论:成功表达了Gankyrin蛋白并制备了其特异性抗体.

PTC组织中MTA1、Gankyrin表达的变化与术后复发、转移的关系

目的探讨甲状腺乳头状癌(PTC)组织中肿瘤转移相关基因蛋白1(MTA1)、癌性锚蛋白重复序列(Gankyrin)蛋白的表达与肿瘤术后复发、转移的关系。方法选取郑州大学第一附属医院2017年1月至2019年3月手术治疗的173例PTC患者术后标本作为PTC组、另选取同期手术获取的甲状腺良性肿瘤组织160例作为良性病变组;采用免疫组化染色检测两组标本中的MTA1、Gankyrin蛋白表达,分析不同年龄、性别、TNM分期、肿瘤病灶大小、淋巴结转移组织中MTA1、Gankyrin蛋白表达情况,采用Logistic回归分析法分析上述各项指标与PTC术后转移复发的关系。结果PTC组MTA1蛋白、Gankyrin蛋白阳性表达率均高于良性病变组,差异有统计学意义(P<0.05);PTC标本中MTA1蛋白阳性表达在不同TNM分期、是否发生淋巴结转移、是否出现术后复发转移的患者中比较,差异有统计学意义(P<0.05);Gankyrin蛋白阳性表达率在术后是否出现复发转移的PTC组织中比较,差异有统计学意义(P<0.05);Logistic回归分析结果显示,MTA1蛋白、Gankyrin蛋白阳性表达及TNM分期为Ⅲ期+Ⅳ期是在术后出现复发转移的独立危险因素(P<0.05)。结论MTA1表达与PTC恶性程度增加有关,MTA1蛋白、Gankyrin蛋白表达增强可能会增大PTC患者术后复发转移的风险。

G蛋白偶联雌激素受体和Gankyrin蛋白在卵巢子宫内膜异位症中的表达及意义

目的:研究新型雌激素受体G蛋白偶联雌激素受体(G protein-coupled estrogen receptor,GPER)和Gankyrin在卵巢子宫内膜异位症中的表达及其临床病理意义,探讨二者在卵巢子宫内膜异位症发生、发展中的作用。方法:采用免疫组织化学法检测GRER和Gankyrin蛋白在卵巢子宫内膜异位症患者的异位内膜、在位内膜及正常子宫内膜中的表达情况及定位分布,结合临床资料分析其与卵巢子宫内膜异位症分期的关系,并评估两者之间的相关性。结果:GPER蛋白在异位、在位及正常内膜中的阳性表达率分别为63.6%,51.5%,21.2%,异位及在位内膜组的阳性表达显著高于正常内膜组(均P<0.0125),而异位与在位内膜间差异无统计学意义(P>0.0125);Gankyrin蛋白在异位、在位及正常内膜中的阳性表达率分别为69.7%,36.4%,9.1%,组间两两比较差异均有统计学意义(均P<0.0125)。在位内膜中,GPER蛋白表达在增生期高于分泌期(P<0.05),Gankyrin蛋白在增生期与分泌期的表达差异无统计学意义(P>0.05)。GPER和Gankyrin蛋白的表达与卵巢子宫内膜异位症的临床分期有关,二者在III^IV期的阳性表达率高于I^II期(均P<0.05)。GPER和Gankyrin蛋白在异位内膜中的表达呈正相关(rs=0.640,P<0.01)。结论:GPER和Gankyrin的表达与卵巢子宫内膜异位症的发生、发展及临床分期有关,二者可能通过相互作用促进异位病灶的形成。

Inhibition of apoptosis by oncogenic hepatitis B virus X protein: Implications for the treatment of hepatocellular carcinoma

Hepatitis B virus X protein(HBx) plays an important role in the development of hepatocellular carcinoma(HCC). In addition, hepatoma upregulated protein(HURP) is a cellular oncogene that is upregulated in a majority of HCC cases. We highlight here recent findings demonstrating a link between HBx, HURP and anti-apoptosis effects observed in cisplatin-treated HCC cells. We observed that Hep3B cells overexpressing HBx display increased HURP mRNA and protein levels, and show resistance to cisplatin-induced apoptosis. Knockdown of HURP in HBx-expressing cells reverses this effect, and sensitizes cells to cisplatin. The anti-apoptotic effect of HBx requires activation of the p38/MAPK pathway as well as expression of SATB1, survivin and HURP. Furthermore, silencing of HURP using short-hairpin RNA promotes accumulation of p53 and reduces cell proliferation in SK-Hep-1 cells(p53^(+/–)), whereas these effects are not observed in p53-mutant Mahlavu cells. Similarly, HURP silencing does not affect the proliferation of H1299 lung carcinoma cells or Hep3 B HCC cells which lack p53. Silencing of HURP sensitizes SK-Hep-1 cells to cisplatin. While HURP overexpression promotes p53 ubiquitination and degradation by the proteasome, HURP silencing reverses these effects. Inoculation of SK-Hep-1 cancer cells in which HURP has been silenced produces smaller tumors than control in nude mice. Besides, gankyrin, a positive regulator of the E3 ubiquitin ligase MDM2, is upregulated following HURP expression, and silencing of gankyrin reduces HURP-mediated downregulation of p53. In addition, we observed a positive correlation between HURP and gankyrin protein levels in HCC patients(r^2 = 0.778; n = 9). These findings suggest a role for the viral protein HBx and the host protein HURP in preventing p53-mediated apoptosis during cancer progression and establishment of chemoresistance.

Gankyrin: A potential target for drug therapy against hepatocellular carcinoma

Hepatocellular Carcinoma is a primary malignant tumor of the liver and gankyrin is an oncoprotein over-expressed in hepatocellular carcinoma. It has been found that Gankyrin protein reduces the level of p53 protein by increasing its ubiquitylation and degradation, following a MDM-2 mediated pathway. Interaction of gankyrin with MDM2 enhances the ubiquitylation of p53. Independent study of this protein molecule revealed that it is identical to the p28 subunit of the 26S proteasome, having seven similar alpha helical ankyrin repeats. Gankyrin also binds to the Tumor Suppressor Protein (TSP) Retinoblastoma (RB), thereby accelerating its phosphorylation and proteasomal degradation. Blocking the expression of Gankyrin with MDM2 in cases of Hepatocellular Carcinoma (HCC) promoted apoptosis in cancer cells. Hence, Gankyrin can be used as a potential target for drug therapy against Hepatocellular Carcinoma.

口腔鳞状细胞癌组织癌性锚蛋白重复序列蛋白的表达及意义

目的探讨癌性锚蛋白重复序列(gann ankyrin repeats,gankyrin)蛋白在口腔鳞状细胞癌中的表达及临床意义。方法采用免疫组化方法检测55例口腔鳞状细胞癌组织及癌旁黏膜组织中gankyrin蛋白表达水平,观察口腔鳞状细胞癌不同临床病理特征下gankyrin蛋白的表达差异,分析其可能的临床价值。结果口腔鳞状细胞癌组织中gankyrin蛋白表达远高于癌旁黏膜组织(P=0.000)。高、中低分化鳞癌组间gankyrin蛋白表达差异有统计学意义(P=0.006);不同性别、年龄及有无淋巴结转移鳞癌组gankyrin蛋白表达差异无统计学意义。结论 Gankyrin蛋白高表达于口腔鳞状细胞癌,且与肿瘤分化程度相关。

Gankyrin在膀胱癌中的表达及临床意义

目的：探讨Gankyrin在膀胱癌中的表达及临床意义。方法：应用免疫组织化学检测2002年9月~2008年10月手术切除的60例膀胱癌组织及癌旁组织中Gankryin的表达水平;应用RT-PCR检测32例新鲜膀胱癌组织中Gankyrin的表达情况,并用统计学方法分析Gankyrin与膀胱癌的TNM分期及分化程度等临床病理资料及预后的关系。结果：Gankyrin在膀胱癌组织中的表达率为73.3%,显著高于癌旁组织的21.7%,且Gankyrin的表达与肿瘤TNM分期及分化程度以及患者预后显著相关（P〈0.05）,而与患者的性别及年龄无关（P〉0.05）。结论：Gankyrin在膀胱癌中普遍表达且与其恶性程度及预后显著相关,可作为判断膀胱癌生物学行为及预后的有效指标。