乳腺癌中PHF2的表达及其临床意义

目的探讨PHF2在乳腺癌细胞和组织中的表达及其与乳腺癌临床病理特征、预后的关系。方法采用q RT-PCR法检测乳腺癌及正常乳腺上皮细胞株(n=7)及配对的癌旁组织(n=30)中PHF2 m RNA的表达水平;应用Western blot法检测乳腺癌细胞株中PHF2蛋白的表达水平,免疫组化法检测乳腺癌组织(n=50)和癌旁组织(n=15)中PHF2蛋白的表达,分析PHF2表达与乳腺癌临床病理特征的关系及预后的关系。结果与正常人乳腺上皮细胞相比,PHF2 m RNA和蛋白在乳腺癌细胞株中表达下调(P0.01);与癌旁组织相比,PHF2 m RNA在90%(27/30)的乳腺癌组织中表达下调(P0.05);免疫组化结果显示:在70%(35/50)的乳腺癌组织中PHF2蛋白表达下调(P0.05);PHF2表达下调与乳腺癌患者的淋巴结转移、ER阴性及预后密切相关(P0.05)。结论 PHF2在乳腺癌中表达下调,可以作为乳腺癌预后判断的指标之一,有望成为乳腺癌治疗的新靶点。     

ACTL6A在宫颈癌中的表达及意义

目的:探讨ACTL6A在宫颈癌中的表达及意义。方法:收集2012年1月至2015年1月间我院收治的93例宫颈癌患者的癌组织和癌旁组织石蜡标本,采用免疫组化法测定ACTL6A表达情况,分析其与临床病理参数及预后之间的关系。应用多因素Cox回归分析预测不良事件的指标。结果:ACTL6A在宫颈癌癌组织阳性表达率(63.5%)明显高于癌旁组织(27.9%)(P<0.05)。ACTL6A蛋白高表达与宫颈癌的淋巴结转移、组织学分解、TNM分期相关,而与患者年龄、肿瘤大小无关。随访时间3-60个月,宫颈癌癌组织高表达ACTL6A蛋白的患者总生存明显低于低表达的患者(P<0.05);其中淋巴结转移、组织学分级、TNM分期和ACTL6A均是宫颈癌患者预后的独立影响指标。结论:ACTL6A与宫颈癌的发生和进展关系密切,是判断宫颈癌患者预后的重要指标。     

乳腺癌患者肌动蛋白样蛋白8表达水平及其与临床病理的关系分析

目的探讨肌动蛋白样蛋白8(ACTL8)在乳腺癌患者中表达水平及其与临床病理的关系。方法选取2010年2月至2015年2月在我院诊治的女性乳腺癌患者(n=100),并随机抽取同时间段内体检的正常女性(n=50),分别取两组女性乳腺组织,进行免疫组织化学染色,观察两组乳腺癌患者ACTL8水平,分析乳腺癌患者ACTL8表达与临床病理关系。结果 ACTL8在正常乳腺组织中阳性例数明显低于乳腺癌组织(P0.05);100例乳腺癌患者中,ACTL8表达阴性42例(42.00%),阳性58例(58.00),其中不同表达患者年龄无差异(P0.05);淋巴结转移、肿瘤大小、临床分期、SBR分级有差异(P0.05)。结论乳腺癌患者ACTL8表达水平较正常女性高,且ACTL8表达与临床病理存在一定关联。     

VEGF、HIF-1α在乳腺癌患者组织中的表达及临床意义

目的:分析血管内皮生长因子(Vascular endothelial growth factor,VEGF)、缺氧诱导因子-1α(Hypoxia inducible factor-1 α,HIF-1α)在乳腺癌患者组织中的表达及临床意义.方法:收集2018年9月至2021年3月于本院经病理组织确诊的78例乳腺癌患者组织标本及其癌旁正常乳腺组织标本资料.采用免疫组织化学染色法检测病灶组织及癌旁正常乳腺组织中VEGF、HIF-1α蛋白表达情况,记录患者临床特征.对比乳腺癌患者的病灶组织与癌旁正常乳腺组织中VEGF、HIF-1α蛋白阳性表达率;分析对比不同临床特征的乳腺癌患者病灶组织中VEGF、HIF-1α蛋白阳性表达情况;病灶组织中VEGF、HIF-1 α蛋白阳性表达对乳腺癌患者淋巴结转移的评估价值.结果:病灶组织中VEGF、HIF-1α蛋白阳性表达率均高于癌旁正常乳腺组织(P＜0.05);有淋巴结转移的患者病灶组织中VEGF、HIF-1α蛋白阳性表达率高于无淋巴结转移的患者(P＜0.05);病灶组织中VEGF、HIF-1α蛋白对乳腺癌患者淋巴结转移评估价值的AUC分别为0.723、0.727,均有一定评估价值.结论:VEGF、HIF-1α蛋白在乳腺癌组织中阳性表达率较高,检测病灶组织中VEGF、HIF-1α蛋白阳性表达有助于评估患者淋巴结转移情况     

乳腺癌组织中着丝粒蛋白CENP-B的表达及意义

目的检测乳腺癌组织中着丝粒蛋白CENP-B的表达,探讨CENP-B的表达与乳腺癌患者临床病理特征的关系及意义。方法分别用RT-PCR、Q-PCR和Western blot法检测4例乳腺癌活检组织及相应癌旁组织中CENP-B mRNA和蛋白表达水平的差异;采用免疫组化SP法检测CENP-B在53例乳腺癌石蜡组织切片中的表达,并采用SPSS 10.0软件分析其与乳腺癌的临床病理特征的关系。结果 CENP-B mRNA和蛋白在4例乳腺癌组织中的表达高于癌旁组织;53例乳腺癌组织中CENP-B阳性率为90.6%(48/53),强阳性率为52.8%(28/53),CENP-B在乳腺肿瘤组织中的表达水平高于癌旁组织。统计分析结果发现CENP-B强阳性与临床分期、T分期密切相关。结论 CENP-B可能成为评价乳腺癌发生、发展的重要生物学指标。     

结直肠腺癌中TKT的表达及临床意义

目的探讨结直肠腺癌中转酮醇酶(transketolase, TKT)的表达及与临床病理特征的关系。方法通过Oncomine和GEO数据库分析TKT基因在结直肠腺癌和癌旁组织中的表达。收集配对结直肠腺癌及癌旁正常肠黏膜组织,分别采用qRT-PCR、Western blot法检测TKT mRNA和蛋白表达,运用免疫组化法检测TKT蛋白表达,并分析其与临床病理特征的相关性。结果 qRT-PCR结果显示:与癌旁组织相比,27例结直肠腺癌组织中TKT mRNA表达显著升高(P<0.001);Western blot结果显示:8例结直肠腺癌组织中TKT蛋白表达高于癌旁组织,在常见结直肠癌细胞株中TKT蛋白表达高于FHC细胞株;两种检测与数据库样本分析一致。免疫组化检测结果显示:92例结直肠腺癌组织中TKT高表达率显著高于癌旁组织(P<0.001),TKT高表达与肿瘤直径(P=0.003)、脉管侵犯(P=0.021)、淋巴结转移(P=0.005)及T分期(P=0.007)相关。结论 TKT在结直肠腺癌中呈高表达,与肿瘤侵袭转移临床病理特征密切相关,其可能参与结直肠腺癌的发生、发展,TKT有望成为结直肠癌预后标志物或潜在的治疗靶点。     

Endoglin在非小细胞肺癌中的表达及临床意义

目的 研究不同转移潜能非小细胞肺癌(NSCLC)细胞系中以及癌与癌旁组织中endoglin (ENG)的表达情况,探讨ENG在NSCLC的发生发展和转移复发中的作用.方法 体外培养5株NSCLC细胞及1株正常支气管上皮细胞(HBE),随机选取22例NSCLC手术切除后癌与癌旁组织样本,分别采用real time PCR及Western blot法检测其中ENG mRNA和蛋白表达情况.采用x2检验分析ENG的表达和临床资料之间的相关性.结果 高转移性3株肺癌细胞系中ENG mRNA和蛋白表达显著升高,而正常细胞株及低转移性2株肺癌细胞中ENG表达缺失.22例肺癌样本中,其中19例癌组织中ENG mRNA和蛋白呈阳性表达(86.36％),且表达量明显高于相对应的癌旁组织(P＜0.01).ENG表达与患者临床病理特征年龄、性别、肿瘤大小、临床分期、病理分级、组织类型等均无关,而与患者淋巴结转移情况密切相关(P＜0.01).结论 ENG在NSCLC中的表达与淋巴结转移显著相关,有可能作为预测NSCLC转移及预后的分子标志.     

乳腺癌和癌旁乳腺组织中Notch1基因mRNA及蛋白的表达

目的 检测Notch1基因mRNA及Notch1蛋白在人乳腺癌和癌旁乳腺组织中的表达,分析其临床病理学意义.方法 应用逆转录聚合酶链反应(RT-PCR)方法榆测60例乳腺浸润性导管癌和60例癌旁乳腺组织中Notch1基因mRNA,应用免疫组织化学SP法检测60例乳腺浸润性导管癌、30例导管原位癌及60例癌旁乳腺组织Notch1蛋白的表达,分析Notch1表达水平与乳腺癌临床病理特征的相关性.结果 Notch1基因mRNA在人乳腺浸润性导管癌及癌旁乳腺组织中均有表达.Notch1蛋白在癌旁乳腺组织和导管原位癌中的阳性牢分别为55%(33/60)、70%(21/30),二者差异无统计学意义(P>0.05),在乳腺浸润性导管癌中的阳性率为90%(54/60),明显高于癌旁乳腺组织和导管原位癌的阳性率(P<0.05).乳腺浸润性导管癌Notch1蛋白的高表达与肿瘤的淋巴结转移(P=0.006)、病理学分级(P=0.001)和TNM分期(P=0.022)均呈显著正相关.结论 乳腺浸润性导管癌存在Notch1蛋白的高表达.Notch1蛋白高表达与乳腺癌的恶变演进有关.Notch1基因的表达可能影响乳腺癌的发生、发展.     

乳腺癌组织及细胞系中CCDC34的表达及其生物学功能

目的 探索乳腺癌组织及细胞系中CCDC34的表达情况及其对乳腺癌细胞增殖能力的影响。方法免疫组织化学染色方法检测34例乳腺癌组织及15例癌旁正常乳腺组织中CCDC34的表达和定位情况;利用Kaplan-Meier Plotter和The Human Protein Atlas数据库分析CCDC34与乳腺癌患者预后的相关性;Western blot检测人正常乳腺细胞MCF-10A及乳腺癌细胞SKBR-3、MCF-7、MDA-MB-453和MDA-MB-231中CCDC34的表达情况;在MCF-7和MDA-MB-231细胞中瞬时转染CCDC34,使用MTT的方法检测CCDC34对乳腺癌细胞增殖能力的影响。结果 CCDC34在乳腺癌组织的表达显著低于癌旁正常乳腺组织（P<0.01）;CCDC34在乳腺癌中的表达水平与患者的不良预后显著负相关（P<0.01）;CCDC34在乳腺癌细胞中的表达普遍低于人正常乳腺细胞MCF-10A(P<0.01);MTT检测结果显示,转染CCDC34显著抑制了乳腺癌细胞MCF-7(Luminal A)和MDA-MB-231(basal)细胞的增殖能力（P<0.05）。结论 乳腺癌组织及细胞系中CCDC34的表达普遍降低,CCDC34能够抑制乳腺癌细胞的增殖能力,并与乳腺癌患者的良好预后相关。     

乳腺癌中RNF2的表达及其临床意义

目的 观察RNF2在乳腺病变组织及乳腺上皮细胞系中的表达,探讨其表达与乳腺癌临床病理特征的关系。方法 运用免疫组化En Vision两步法检测21例乳腺良性病变组织和112例乳腺癌组织中RNF2蛋白的表达,分析其表达与乳腺癌临床病理特征的关系。应用qRT-PCR法检测2株乳腺正常上皮细胞系和5株乳腺癌细胞系中RNF2基因的表达。结果 乳腺癌组织中RNF2蛋白表达水平与乳腺良性病变组织相比显著升高(P0.05),且其表达与乳腺癌肿瘤直径、腋窝淋巴结转移数目、TNM分期显著相关(P均0.05);与患者年龄、组织学分级、ER、PR、HER-2表达情况无显著相关(P均0.05);乳腺癌细胞系RNF2 mRNA水平亦高于乳腺良性上皮细胞系(P0.05)。结论 RNF2在乳腺癌组织中高表达,可作为乳腺癌治疗的潜在靶点。     

Expression of flotillin-2 in human non-small cell lung cancer and its correlation with tumor progression and patient survival

Introduction: Recent studies have revealed that flotillin-2 (FLOT2) played important roles in cancer progression. The aim of this study was to investigate the clinicopathologic and prognostic significance of FLOT2 expression in human non-small cell lung cancer (NSCLC). Methods: Quantitative real-time PCR (qRT-PCR) was performed to detect FLOT2 mRNA expression in lung cancer cell lines, normal bronchial epithelial cells, 24 pairs of NSCLC tissues and matched adjacent non-tumor tissues. Immunohistochemistry (IHC) was performed to examine FLOT2 protein expression in paraffin-embedded tissues from 90 NSCLC patients. Statistical analyses were performed to evaluate the clinicopathological significance of FLOT2 expression. Results: FLOT2 mRNA expression was evidently up-regulated in lung cancer cell lines and NSCLC tissues compared with normal bronchial epithelial cells and adjacent non-tumor tissues. In the 90 cases of tested NSCLC samples, FLOT2 protein level was positively correlated with tumor stage, and lymph node metastasis. Patients with high FLOT2 expression had shorter overall survival compared with the low FLOT2 expression group. Univariate and multivariate analyses indicated that high FLOT2 expression was an independent poor prognostic factor for NSCLC patients. Conclusions: Our findings provided that high FLOT2 expression was associated with poor outcomes in NSCLC patients, and FLOT2 could be a potential prognostic biomarker for lung cancer progression.     

Overexpression of histone demethylase JMJD5 promotes metastasis and indicates a poor prognosis in breast cancer

In this study, we showed the expression of JMJD5 was increased in breast cancer tissues and breast adenocarcinoma cell lines MCF-7 as well as triple negative breast cancer cell lines MDA-MB-231 compared with paired adjacent normal mammary tissues and normal mammary epithelial cell lines MCF-10A. The higher expression of JMJD5 was significantly corresponded with clinical stage, histological grade and lymph node metastasis. Overexpression of JMJD5 promoted cell invasion and induce EMT, while JMJD5 siRNA inhibits MDA-MB-231 cells invasion in vitro. Moreover, qChIP analysis revealed the Snail family proteins Snai1 was the direct target of JMJD5 in breast cancer cells. Luciferase reporter assays suggested that the overexpression of JMJD5 resulted in the activation of Snail1 promoter-driven luciferase reporter. The changes in the level of RNA and protein implied that the activation of Snail was the important mechanisms by which JMJD5 triggers metastasis. We also detected the higher expression of JMJD5 protein was an independent unfavorable biomarker for worse overall survival in breast cancer patients. Therefore, our results identified an important role for JMJD5 in breast cancer through the regulation of snail1.     

HPD overexpression predicts poor prognosis in breast cancer

BackgroundThe enzyme, 4-hydroxyphenylpyruvate dioxygenase (HPD), is critical to tyrosine metabolism; its deficiency can cause tyrosinemia. However, its precise contribution to tumorigenesis is unclear. Here, we investigated the correlation between HPD expression and prognosis in patients with breast cancer.Methods145 breast cancer specimens were selected to analyze HPD protein expression by immunohistochemistry and evaluate its relationship to patients’ clinicopathological features. HPD localization was confirmed in MCF-7 and MDA-MB-231 breast cancer cells, using immunofluorescence staining. The expression of HPD protein was detected in breast cancer and cancer-adjacent normal tissues using Western blot analysis. Survival rates were calculated by the Kaplan–Meier method.ResultsWe found that HPD protein was mainly located in the cytoplasm/nucleoli/perinucleus in breast cancer cells, as shown by immunofluorescence staining in MCF-7 and MDA-MB-231 cells, and immunohistochemistry in breast cancer and adjacent normal tissues (HPD protein expression—breast cancer: 46.9% [68/145], ductal carcinoma in situ [DCIS]: 22.6% [12/53], and normal tissues: only 4.8% [2/42]). Similarly, the Western blot results further confirmed the increased expression of HPD in breast cancer compared with cancer-adjacent normal tissues (P < 0.05). HPD expression level was positively correlated with histological grade and clinical stage, and inversely correlated with 10-year overall survival (OS) rates, in patients with breast cancer. Among patients with breast cancer, those with high HPD expression had worse OS rates than those with low HPD expression. Additionally, when patients were subgrouped by disease stage or grade, those with high HPD expression had worse OS rates than those with low HPD expression for each respective stage or grade.ConclusionsOur findings indicate that HPD may be a useful prognostic predictor, and a potential therapeutic target for patients with breast cancer.     

WNT5B过表达对人乳腺癌MCF-7细胞活力及凋亡的影响

目的:检测WNT5B在人正常乳腺上皮细胞和乳腺癌细胞株中的表达,及过表达WNT5B后对人乳腺癌MCF-7细胞活力及凋亡的影响,探讨WNT5B在乳腺癌发生发展中的作用。方法:采用RT-PCR法检测正常乳腺上皮细胞和乳腺癌细胞中WNT5B的mRNA表达水平,将WNT5B的表达载体pc DNA3.1/WNT5B和空白对照载体pc DNA3.1分别瞬时转染人乳腺癌MCF-7细胞,通过real-time PCR和Western blotting法分别在mRNA和蛋白水平验证WNT5B的表达效率;并通过CCK-8的方法检测WNT5B过表达对人乳腺癌MCF-7细胞活力的影响,通过流式细胞术分别检测过表达WNT5B对MCF-7细胞周期分布及凋亡比例的影响。结果:与正常乳腺上皮细胞相比,WNT5B在乳腺癌细胞中低表达;在乳腺癌细胞MCF-7中转染WNT5B表达质粒后,WNT5B表达上调,差异有统计学显著性(P0.05);与对照组相比,过表达WNT5B后,MCF7细胞的活力明显降低,处于S期细胞的量明显增加,G1和G2/M期细胞明显减少,且细胞凋亡比例明显增加,差异均有统计学显著性(P0.05)。结论:WNT5B在乳腺癌细胞中低表达,过表达WNT5B可使人乳腺癌细胞MCF-7阻滞在S期,抑制细胞增殖,促进细胞凋亡。     

婆罗双树样基因4在人前列腺癌细胞和组织中的表达及其与临床预后之间的关系

 目的：评估婆罗双树样基因4（SALL4）在人前列腺癌细胞系和前列腺癌组织中的表达情况,并明确其表达水平和临床病理学参数间的关系。方法：用免疫荧光技术、RT-PCR和Western blotting检测SALL4在LNCaP、DU145、PC-3细胞系和RWPE-1正常前列腺上皮细胞系中的表达。同时用免疫组化方法检测SALL4在前列腺增生及前列腺癌组织中的表达,并探讨其与Gleason评分等临床病理参数的关系。结果： SALL4蛋白在细胞中主要表达于胞浆,在3种前列腺癌细胞株中SALL4蛋白表达水平要明显高于正常前列腺上皮细胞RWPE-1（P＜005）,而SALL4 mRNA表达水平在4种细胞系中无明显差异（P＞0.05）。免疫组化结果显示SALL4在前列腺癌组织中的表达水平明显高于增生和正常前列腺组织（P＜0.01）。SALL4蛋白表达水平与Gleason评分、前列腺癌临床分期、预后及组织前列腺特异抗原（PSA）表达密切相关,而与患者年龄、治疗前血清总PSA水平、前列腺体积及组织雄激素受体表达无明显相关性。结论：SALL4蛋白在前列腺癌中高表达,提示其在前列腺癌的发生、发展过程中可能起重要作用,有可能成为诊断前列腺癌新的肿瘤标志物及评估其恶性程度、进展和预后的参考指标。     

Detection of laminin receptor mRNA in human cancer cell lines and colorectal tissues by in situ hybridization.

The 67-kd high-affinity laminin receptor (67 LR) is a gene product whose expression appears to be associated with the invasive and metastatic phenotype of a variety of human cancer cells. Northern blot hybridization has been routinely used to quantify the level of 67 LR mRNA from total cellular RNA extracts of homogenized tissue specimens or in vitro grown cell populations. This technique is useful to assess the average expression of the 67 LR mRNA of a particular sample but does not provide information about expression in specific cell types nor about heterogeneity of expression from cell to cell. In this study, we analyzed the expression of 67 LR mRNA in four human cancer cell lines with varying degrees of expression of 67 LR protein (renal cancer A-704, breast carcinoma MCF-7/4 and MCF-7/7, and pancreatic cancer Panc-1) using in situ hybridization performed with 67 LR riboprobes. Total cellular RNA was simultaneously extracted from the cell lines and hybridized on Northern blots with a 67 LR cDNA probe to assess the validity of the mRNA detection by in situ hybridization. Sixty-seven LR mRNA expression was higher in Panc-1 and MCF-7/4 cells than in MCF-7/7 and renal carcinoma A-704. There was a direct correlation (R2 = 0.88) between the in situ hybridization analysis and the mRNA levels detected by Northern blot analysis. The in situ hybridization method showed a heterogeneous expression of the 67 LR mRNA in the four cell lines with different subpopulations of cells showing a range from negative to high levels of the message. Sixteen freshly frozen human colorectal tissues (seven adenocarcinomas, five matched normal mucosae, and four adenomas) were also analyzed by in situ hybridization. The 67 LR mRNA was localized in normal and neoplastic epithelial cells. Adenocarcinoma cells showed a 1.6- to 5-fold higher expression (P < 0.02 according to the Wilcoxon-Mann-Whitney test) than did epithelial colonic cells from normal mucosae or adenomas. The signal tended to be stronger in poorly differentiated carcinomas and carcinomas with metastases than in moderately differentiated and nonmetastatic tumors. We conclude that the high expression of 67 LR mRNA in colorectal tumors is due to an increased production by tumor cells. Furthermore, in situ hybridization is an effective method to detect the expression of LR mRNA in cultured cell lines as well as in frozen tissue sections.