miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。     

Twist expression associated with the epithelial-mesenchymal transition in gastric cancer

This study aimed to investigate the expression of Twist in gastric cancer tissues and its correlation between Twist and the epithelial-mesenchymal transition (EMT). By means of RT-PCR and Western blot, the mRNA and protein expressions of Twist, E-cadherin, and Vimentin in 61 gastric cancer tissues and adjacent normal tissues were detected. The positive rates of Twist, E-cadherin, and Vimentin mRNA expression in gastric cancer tissues were 73.9. 40.6, and 60.9 %, respectively; compared to the expression of these genes in adjacent normal tissues (2.9, 75.4, and 27.5 %), the differences were significant (p < 0.05). The E-cadherin protein expression level in gastric cancer tissues was significantly lower than that in the adjacent normal tissues (p < 0.05). After the transfection of Twist siRNA into the MKN45 cells, the protein expression of Twist was significantly reduced (p < 0.05), the protein expression of E-cadherin was significantly increased, and the number of cells that passed through the Transwell chamber was significantly lower than that in the non-transfected control group as well as the transfected control group (p < 0.05). Twist may be associated with the epithelial-mesenchymal transition in gastric cancer and the tumorigenesis, invasion, and metastasis of gastric cancer.     

MT1F mRNA在结肠癌组织中的表达及其临床病理意义

目的:探讨MT1F mRNA在结肠癌组织中的表达及其临床病理意义。方法:采用Taq Man探针实时荧光定量Real-time PCR检测40例结肠癌及对应正常粘膜组织中MT1F mRNA的表达,并通过免疫组化检测Metallothionein(MT)的蛋白表达。结果:82.5%(33/40例)的结肠癌组织MT1F mRNA表达较对应正常组织明显下调,降低2.4倍至113倍不等。MT1F mRNA水平与结肠癌患者的年龄、性别、肿瘤部位、肉眼形态、直径、分化及Dukes分期无关。在MT1F mRNA低表达的病例中,癌组织MT蛋白表达低于对应正常组织(P0.05)。结论:结肠癌组织MT1F mRNA水平显著下调,但与结肠癌的临床病理特征无关。     

Survivin 及Anx-A1 在肝癌组织中的表达及其临床意义

目的:探讨Survivin及Anx-A1在肝癌组织中的表达及其临床意义。方法:收集原发性肝癌病例45例,采用免疫组织化染色法检测Survivin及Anx-A1在肝癌组织及癌旁正常组织中的表达,分析Survivin及Anx-A1的表达与肝癌临床病理特征的关系。结果:Survivin在肝癌组织中的阳性表达率为86.67%,在癌旁正常组织中的阳性表达率为17.78%;Anx-A1在肝癌组织中的阳性表达率为46.67%,在癌旁正常组织中的阳性表达率为8.89%;Survivin及Anx-A1在肝癌组织中的阳性表达率均显著高于癌旁正常组织,差异具有统计学意义(P0.05);不同肿瘤分级患者肝癌组织中Survivin与Anx-A1的表达水平存在显著差异(P0.01),肿瘤分级越高,Survivin与Anx-A1表达水平越高。结论:Survivin及Anx-A1的表达与肝癌的发生发展密切相关,可用于肝癌的辅助诊断。     

Glutathione and GSH-dependent enzymes in patients with liver cirrhosis and hepatocellular carcinoma

We investigated glutathione level, activities of selenium independent GSH peroxidase, selenium dependent GSH peroxidase, GSH S-transferase, GSH reductase and the rate of lipid peroxidation expressed as the level of malondialdehyde in liver tissues obtained from patients diagnosed with cirrhosis or hepatocellular carcinoma. GSH level was found to be lower in malignant tissues compared to adjacent normal tissues and it was higher in cancer than in cirrhotic tissue. Non-Se-GSH-Px activity was lower in cancer tissue compared with adjacent normal liver or cirrhotic tissue, while Se-GSH-Px activity in cancer was found to be similar to its activity in cirrhotic tissue and lower compared to control tissue. An increase in GST activity was observed in cirrhotic tissue compared with cancer tissue, whereas the GST activity in cancer was lower than in adjacent normal tissue. The activity of GSH-R was similar in cirrhotic and cancer tissues, but higher in cancer tissue compared to control liver tissue. An increased level of MDA was found in cancer tissue in comparison with control tissue, besides its level was higher in cancer tissue than in cirrhotic tissue. Our results show that the antioxidant system of cirrhosis and hepatocellular carcinoma is severely impaired. This is associated with changes of glutathione level and activities of GSH-dependent enzymes in liver tissue. GSH and enzymes cooperating with it are important factors in the process of liver diseases development.     

Cx43 与Smo 基因在胰腺癌中的表达及其临床意义

目的:研究细胞间隙连接蛋白43(Cx43)与信号通路基因Smoothened(Smo)在胰腺癌中的表达及其临床意义。方法:选择从2013年1月~2015年12月在医院接受手术治疗的胰腺癌患者53例切除组织及与癌组织相配对的3cm外癌旁组织,对比不同胰腺组织中Cx43与Smo阳性表达率,及Cx43 m RNA与Smo m RNA表达水平,分析胰腺癌组织中Cx43和Smo表达与胰腺癌的病理特征之间的关系及两者的相关性。结果:胰腺癌组织中Cx43的阳性表达率及Cx43 m RNA表达水平明显低于在癌旁组织,而Smo的阳性表达率及Smo m RNA表达水平明显高于癌旁组织,差异均有统计学意义(均P0.05)。胰腺癌患者组织学分级为Ⅲ级、有淋巴结转移者的Cx43 m RNA表达水平分别明显低于Ⅰ~Ⅱ级、无淋巴结转移者,Smo m RNA表达水平则明显高于Ⅰ~Ⅱ级、无淋巴结转移者,差异均有统计学意义(均P0.05)。Pearson相关性分析结果发现,胰腺癌组织中Cx43 m RNA和Smo m RNA表达呈负相关关系(r=-0.846,P=0.000)。结论:Cx43 m RNA在胰腺癌中的表达下降,而Smo m RNA则表达上调,临床监测Cx43及Smo基因的表达情况,有助于评价患者的病情与预后。     

Clinical relevance of cyclooxygenase-2 and matrix metalloproteinases (MMP-2 and MT1-MMP) in human breast cancer tissue

Breast cancer (BC) is the most common neoplasm among women in most developed countries, including Egypt. Elevated levels of certain proteins in human BC are associated with unfavorable prognosis and progressive stages of the disease. The aim of our study was to evaluate the protein expression profile and prognostic significance of cyclooxygenase-2 (COX-2), matrix metalloproteinase-2 (MMP-2), MMP-9 and membrane type 1-MMP (MT1-MMP) and their interaction in operable BC patients. The protein expression of COX-2, MMP-2 and MT1-MMP were evaluated by western blot technique, whereas enzymatic activity of MMP-2 and MMP-9 was determined by zymography in 47 breast cancer patients as well as normal adjacent tissues. Also, the correlation between these proteins and age, tumor size, LN stage, TNM stage, estrogen receptor, progesterone receptor, disease-free survival, and overall survival (OS) has been investigated. As compared to adjacent normal tissues, COX-2, MMP-2 and MT1-MMP were over-expressed in 43, 64, and 60 % of tumor tissues, respectively. In the same pattern, the activity of MMP-2 (62 %) and MMP-9 (45 %) was elevated in BC tissues. Multivariate analysis showed a positive correlation between the protein expression of COX-2, MMP-2, and MT1-MMP and the activity of MMP-2 and MMP-9 in BC patients. However, the enzymatic activity showed no correlation with clinicopathological features. This study confirms the preclinical evidence that COX-2 increased the expression of MT1-MMP, which in turn activates MMP-2. The lack of correlation with clinicopathological features, OS or disease-free survival ascertains the complexity of tumor progression and metastasis with many pro- and counter regulatory factors.     

基质金属蛋白酶及其抑制剂在乳腺癌中的表达

目的:研究基质金属蛋白酶2(Matrix Metalloproteinase-2,MMP-2),基质金属蛋白酶7(MMP-7),基质金属蛋白酶9(MMP-9),膜型基质金属蛋白酶(Membrane Type-1 Matrix Metalloproteinase,MT1-MMP),金属蛋白酶组织抑制剂1(Tissue Inhibitor of Metalloproteinase,TIMP-1),金属蛋白酶组织抑制剂2(TIMP-2)在乳腺癌组织中mRNA的表达,及与临床病理变量之间的关联。方法:采用150例乳腺癌患者的组织样本。使用半定量逆转录-聚合酶链反应(RT-PCR)法来测定肿瘤组织和正常乳腺组织中MMP-2,MMP-7,MMP-9,MT1-MMP,TIMP-1和TIMP-2的mRNA表达。结果:MMP-2,MMP-7,MMP-9,MT1-MMP,TIMP-1和TIMP-2在乳腺癌中的mRNA表达显著高于正常组织。结论:MMP-2,MMP-7,MMP-9,和MTI-MMP的表达增加和临床病理参数之间的关联,可以用来预测乳腺癌的侵害行为。     

Increased Expression of Pregnancy Up-Regulated Non-Ubiquitous Calmodulin Kinase Is Associated with Poor Prognosis in Clear Cell Renal Cell Carcinoma

Purpose

The aims of this study were to evaluate the clinical significance and potential prognostic value of pregnancy up-regulated non-ubiquitous calmodulin kinase (PNCK) in clear cell renal cell carcinoma (ccRCC) patients.

Materials and Methods

The expression of PNCK mRNA was determined in 24 paired samples of ccRCCs and adjacent normal tissues using real-time RT-PCR. The expression of PNCK was determined in 248 samples of ccRCCs and 92 paired samples of adjacent normal tissues by immunohistochemical analysis. Statistical analysis was performed to define the relationship between PNCK expression and the clinical features of ccRCC.

Results

The mRNA level of PNCK was significantly higher in tumorous tissues than in the adjacent non-tumorous tissues (p<0.001). An immunohistochemical analysis of 92 paired tissue specimens showed that PNCK expression was higher in tumorous tissues than in the adjacent non-tumorous tissues (p<0.001). Moreover, there was a significant correlation between the PNCK expression and various clinicopathological parameters such as Fuhrman grade (p = 0.011), tumor size (p<0.001), T stage (p<0.001) and N stage (p = 0.015). Patients with higher PNCK expression had shorter overall survival time than those with lower PNCK expression (p<0.001). Multivariate analysis indicated that PNCK expression was an independent predictor for poor survival of ccRCC patients.

Conclusions

To our knowledge, this is the first study that determines the relationship between PNCK and prognosis in ccRCC. We found that increased PNCK expression is associated with poor prognosis in ccRCC. PNCK may represent a novel prognostic marker for ccRCC.     

Osteopontin-Enhanced Hepatic Metastasis of Colorectal Cancer Cells

Liver metastasis is a major cause of mortality from colorectal cancer (CRC). However, mechanisms underlying this process are largely unknown. Osteopontin (OPN) is a secreted phosphorylated glycoprotein that is involved in tumor migration and metastasis. The role of OPN in cancer is currently unclear. In this study, OPN mRNA was examined in tissues from CRC, adjacent normal mucosa, and liver metastatic lesions using quantitative real-time PCR analysis. The protein expression of OPN and its receptors (integrin αv and CD44 v6) was detected by using an immunohistochemical (IHC) method. The role of OPN in liver metastasis was studied in established colon cancer Colo-205 and SW-480 cell lines transfected with sense- or antisense-OPN eukaryotic expression plasmids by flow cytometry and cell adhesion assay. Florescence redistribution after photobleaching (FRAP) was used to study gap functional intercellular communication (GJIC) among OPN-transfected cells. It was found that OPN was highly expressed in metastatic hepatic lesions from CRC compared to primary CRC tissue and adjacent normal mucosa. The expression of OPN mRNA in tumor tissues was significantly related with the CRC stages. OPN expression was also detected in normal hepatocytes surrounding CRC metastatic lesions. Two known receptors of OPN, integrin αv and CD44v6 proteins, were strongly expressed in hepatocytes from normal liver. CRC cells with forced OPN expression exhibited increased heterotypic adhesion with endothelial cells and weakened intercellular communication. OPN plays a significant role in CRC metastasis to liver through interaction with its receptors in hepatocytes, decreased homotypic adhesion, and enhanced heterotypic adhesion.     

Nrf2 和HO_1 在肾癌组织中的表达及意义

目的：探讨肾癌组织Nrf2 和HO_1 表达水平变化及其临床意义。方法：选择经手术切除肾癌组织标本42 例及正常肾组织42 例作为研究对象,42 例患者均未接受化疗或者放疗,采取免疫组织化SABC法测定Nrf2 和HO_1 表达水平,并进行统计学分析。 结果：Nrf2 及HO_1 于肾癌组织及正常肾组织均有不同程度表达,且主要表达于肿瘤细胞胞浆,肾癌组中Nrf2 阳性35 例 （83.3%）,显著高于正常肾组织14例（33.3%）,差异具有统计学意义,P<0.05。肾癌组HO_1 表达阳性31 例（73.8）,显著高于正常 肾组织组15 例（35.7）,差异具有统计学意义,P<0.05。肾癌组织Nrf2和HO_1 表达水平成正相关,r=0.536。结论：肾癌组织Nrf2 和 HO_1表达水平较正常组织更高,且两者之间呈正相关,表明Nrf2 和HO_1 在肾癌进展中起着重要作用。     

乙醛脱氢酶1和肿瘤坏死因子相关诱导凋亡配体在膀胱癌组织中的表达及其临床意义

目的:探讨乙醛脱氢酶1(ALDH-1)和肿瘤坏死因子相关诱导凋亡配体(TRAIL)在膀胱癌组织中的表达及其临床意义。方法:选取2015年3月到2018年1月在河北北方学院附属第一医院进行治疗的膀胱癌患者70例,收集其手术切除的癌组织和癌旁正常组织,采用免疫组化法检测癌组织和癌旁正常组织中ALDH-1、TRAIL表达情况,分析ALDH-1、TRAIL的表达与膀胱癌患者的临床病理特征的关系及癌组织中ALDH-1、TRAIL表达的相关性。结果:癌组织中的ALDH-1的阳性表达率高于癌旁正常组织,TRAIL的阳性表达率低于癌旁正常组织(P0.05)。膀胱癌患者的ALDH-1阳性表达率与年龄、性别、分化程度、肿瘤数量无关(P0.05),临床分期为T2-T3期、有淋巴结转移的膀胱癌患者ALDH-1阳性表达率高于临床分期为Ta-T1期、无淋巴结转移的膀胱癌患者(P0.05)。膀胱癌患者的TRAIL阳性表达率与年龄、性别、临床分期、淋巴结转移、肿瘤数量无关(P0.05),高分化的膀胱癌患者TRAIL阳性表达率高于中低分化的膀胱癌患者(P0.05)。Pearson相关性分析显示,癌组织中ALDH-1、TRAIL表达无明显的相关性(P0.05)。结论:膀胱癌组织中ALDH-1的表达偏高且与临床分期和淋巴结转移有关,TRAIL的表达偏低且与分化程度有关,但ALDH-1和TRAIL之间无相关性,需进一步探讨与研究。     

基质金属蛋白酶和组织金属蛋白酶抑制剂在肾细胞癌组织中的表达

目的:基质金属蛋白酶及组织金属蛋白酶抑制剂在肾细胞癌转移中占有重要的作用,研究肾细胞癌组织中MMP-2、MMP-9、TIMP-1和TIMP-2的表达情况,为肾癌转移的治疗提供理论依据。方法:选取36例肾细胞癌肾组织标本,从相同的肾细胞癌组织及癌旁肾组织获得对照样本,均进行根治性肾切除手术切除。肿瘤分期按TNM分期标准。为了统计评估,肿瘤1期和2期为低级,3期以上为高级。RT-PCR检测肿瘤和正常组织中的MMP-2、MMP-9、TIMP-1和TIMP-2的表达。结果:不同样本MMPs和TIMPs表达水平各不相同。肾细胞癌组织中MMP-2、MMP-9、TIMP-1、TIMP-2在肾细胞癌中的表达明显高于正常肾组织(P0.05)。在肾细胞癌的肿瘤分期方面,MMP-2与MMP-9和肿瘤的分期显著相关,随着肿瘤分期的增加,MMP-2与MMP-9的表达明显升高(P0.05),而TIMP-1与TIMP-2与肿瘤的分期无关。结论:肾细胞癌组织中TIMP-2、MMP-2,MMP-9,TIMP-1的mRNA表达显著高于正常肾组织,抑制MMPS的表达将成为治疗肾细胞癌转移的新的方向。     

Livin蛋白在乳腺癌中的表达及与Bcl-2的关系

目的 探讨凋亡抑制基因Livin和Bel-2蛋白在乳腺癌组织中表达及与临床病理参数的关系。方法应用western blot和免疫组织化学SP法检测90例乳腺癌、30例乳腺良性病变和15例乳腺癌旁组织中Livin和Bcl-2蛋白的表达情况,分析两种蛋白表达的相关性。结果Livin蛋白在乳腺癌组织中的阳性表达率及蛋白表达量明显高于癌旁组织和乳腺良性病变,差异有显著性（P〈0．05）。Livin蛋白在乳腺癌中的阳性表达与肿瘤大小,组织学类型,组织学分级及淋巴结转移无关（P〉0．05）,但随临床分期的增加而升高（P〈0．05）;Livin和Bel-2蛋白在乳腺癌组织中的表达显著相关（P〈0．05）。结论Livin蛋白在乳腺癌组织中表达上调,且与病理分期有关,提示Livin蛋白可通过抑制细胞凋亡促进乳腺癌的发生、发展,Livin与Bel-2蛋白可能在乳腺癌的演进中起着协同作用。     

乳腺癌组织中转化生长因子-beta1 和血管内皮生长因子的表达 及其与血管生成的关系

目的:研究转化生长因子-β1(transforming growth factor-β1,TGF-β1)和血管内皮生长因子(vascular endothelial cell growth factor,VEGF)在乳腺癌组织中的表达及其与血管生成的关系。方法:选取65例手术切除乳腺癌蜡块标本及其周围正常乳腺组织,分为两组:A组为对照组,检测标本为乳腺癌癌旁正常乳腺组织;B组为实验组,检测标本为乳腺癌组织,采用免疫组织化学染色和形态计量检测TGF-β1和VEGF在乳腺癌组织中的表达。利用CD34相关抗原标记血管内皮细胞,计数微血管密度(intratumoral mier oveseulardensity,MVD),并分析其与TGF-β1和VEGF表达的关系。结果:65例乳腺癌组织中,TGF-β1的阳性表达率为69.23%(45/65),TGF-β1阳性表达者MVD值(25.31±4.05)显著高于TGF-β1阴性表达者(21.23±4.29);VEGF的阳性表达率为78.46%(51/65),VEGF阳性表达者MVD值(26.62±3.41)亦明显显著高于VEGF阴性表达者(18.95±6.52)(均P<0.05)。不同病理类型的乳腺癌组织中TGF-β1、VEGF的阳性表达率比较差异无统计学意义(P>0.05),但TGF-β1、VEGF的阳性表达与乳腺癌的组织分级、淋巴结转移呈显著正相关(均P<0.05),且组织学分级越高、淋巴结转移越多,MVD值越大。结论:TGF-β1与VEGF在乳腺癌组织的表达高于正常乳腺组织,并与乳腺癌肿瘤血管的生成有关,二者有望作为乳腺癌恶性程度、浸润转移等生物学行为的评估指标。     

Dysregulation of Claudin family genes in colorectal cancer in a Chinese population

Claudins play an important role in tumor metastasis and in invasiveness of colorectal cancer (CRC). We have evaluated the relationship between CRC and expression of the claudin genes in Chinese patients with CRC. We measured CLDN1 and CLDN7 mRNA using quantitative PCR, and protein levels with immunohistochemistry in cancer tissues and adjacent normal tissue. Cancer tissues had significantly higher levels of CLDN1, and significantly lower levels of CLDN3, CLDN4, and CLDN7 than did normal tissue. CLDN3, CLDN4, and CLDN7 expression levels were higher in CRC of the protruded type than in CRC of the infiltrative type. Expression of CLDN7 correlated with lymph node metastasis. Stage N0 cancer tissues had higher levels of CLDN7 than did stages N1 and N2, suggesting that CLDN7 expression was closely related to the extent of lymph node metastasis. CLDN1 protein was upregulated, but CLDN7 protein was downregulated in cancer tissues when compared with expression in adjacent normal tissues. In conclusion, CLDN3, CLDN4, and CLDN7 were significantly downregulated, whereas CLDN1 was significantly upregulated in CRC. The altered expression of claudin genes may play a role in the initiation and development of CRC.     

甲状腺癌组织中ALCAM 的表达及其与临床病理参数及预后的关系

目的:探讨甲状腺癌组织中活化白细胞黏附分子(activated leukocyte cell adhesion molecule,ALCAM)的表达及其与临床病理参数及预后的关系。方法:选择2011年1月到2013年1月在我院确诊为甲状腺癌的患者60例作为研究对象。收集所有患者甲状腺癌组织及其癌旁正常组织标本,采用免疫组织化学法检测不同组织中ALCAM蛋白表达,分析其与甲状腺癌患者临床病理参数的相关性,并对所有患者随访3年以上,并记录3年生存率。结果:ALCAM蛋白在甲状腺癌组织中的总阳性率为71.67%,明显高于癌旁正常组织的31.67%(P0.05)。ALCAM蛋白在高分化型甲状腺癌组织中的表达率明显高于低分化及无分化型癌组织(P0.05)。ALCAM蛋白高表达的甲状腺癌患者的3年生存率为81.45%,明显高于ALCAM蛋白低表达者的51.32%(P0.05)。结论:组织中ALCAM蛋白表达与甲状腺癌的发生、发展及患者预后密切相关。