lncRNA HULC在膀胱癌组织中的表达及其对5637细胞恶性生物学行为的影响

目的:探讨长链非编码RNA(lncRNA)HULC在膀胱癌组织中的表达及其与患者临床病理特征的关系,以及沉默HULC对膀胱癌5637细胞增殖、凋亡、迁移与侵袭的影响.方法:选取2014年6月至2017年12月郑州人民医院手术切除的102例膀胱癌患者的癌及癌旁组织标本,以及人膀胱癌细胞株5637和人正常膀胱上皮细胞株SV...     

Expression and significance of m1A transmethylase,hTrm6p/hTrm61p and its related gene hTrm6/hTrm61 in bladder urothelial carcinoma

Objective: To discuss the expression of hTrm6p/hTrm61p in bladder urothelial carcinoma tissue and its relationship with m1A level in urine, as well as the influences of hTrm6/hTrm61 on the proliferation and apoptosis of cancer cell line on urothelium. Methods: m1A levels in urine of 32 patients of bladder urothelial carcinoma and normal people were detected by HPLC/ESI-Q-TOF-MS, hTrm6p/hTrm61p expression levels in cancer tissue and para-carcinoma tissue of the same patient were detected by western blotting, and hTrm61 expressions of cancer cell line T24, 5637, and EJ of bladder urothelium and kidney cell line HEK-293 of human embryo were detected by RT-qPCR. The hTrm61 high-expression cell line is selected to detect the situation of proliferation and apoptosis by CCK-8 and flow cytometry (FCM) after knocking out hTrm61 with siRNA. Results: m1A level in urine of carcinoma of urinary bladder is significantly higher than that of normal people, and hTrm6p/hTrm61p expression level in cancer tissue is significantly higher than that in para-carcinoma tissue and has linear correlation with m1A level in urine. The hTrm61 expression in cell line 5637 is significantly higher than that of T24, EJ, and HEK293, and apoptosis is significantly affected after hTrm61 is knocked out from 5637 cell line. Conclusion: High-level expression of hTrm6p/hTrm61p is an important reason for high emission of m1A in urine, and hTrm6/hTrm61 promotes the occurrence of carcinoma of urinary bladder.     

膀胱癌细胞Cx43对顺铂抗肿瘤作用影响研究

目的 连接蛋白43(connexin43,Cx43)在人体正常组织及肿瘤组织中广泛表达,并且参与细胞的生长控制和组织分化.本研究探讨采用RNA干扰技术沉默膀胱癌5637细胞株Cx43蛋白表达对顺铂化疗敏感性的影响.方法 体外培养膀胱癌5637细胞和正常尿路上皮SV-HUC-1细胞,采用蛋白质印迹法检测膀胱癌5637细胞系和正常尿路上皮SV-HUC-1细胞中Cx43蛋白表达,应用免疫荧光技术检测膀胱癌5637细胞系中Cx43蛋白的定位.采用RNA干扰技术沉默膀胱肿瘤5637细胞株Cx43蛋白的表达并用顺铂(3μg/mL)处理SiRNA-Cx43组(实验组)和SiRNA-Control(对照组)后,通过CCK-8检测实验组、对照组及野生型5637细胞的增殖情况;采用流式细胞术检测两组癌细胞的凋亡;采用蛋白质印迹法检测顺铂处理后野生型5637细胞后细胞中Cx43、Cleaved Caspase-3的表达及实验组和对照组中Cx43、Cleaved Caspase-3和Bcl-2的表达.结果 膀胱癌5637细胞中Cx43表达较正常尿路上皮细胞高,相对蛋白表达量分别为1.013±0.102和0.556±0.054,两细胞系比较差异有统计学意义,t=3.789,P=0.019;免疫荧光检测Cx43主要定位于细胞质中.CCK-8结果显示,膀胱癌5637细胞随着顺铂药物的浓度(0.75、1.5、3和6μg/mL)和时间(0、1、2、3 d)的增加,细胞的增殖减少,采用重复测量方差分析,各组间比较,差异有统计学意义,F=153.634,P＜0.001;实验组增殖较对照组明显减少,差异有统计学意义,F=9.949,P=0.02.流式细胞仪检测结果显示,实验组和对照组凋亡率分别为(63.00±4.58)％和(34.33±6.03)％,差异有统计学意义,t=7.457,P＜0.01.蛋白质印迹法结果显示,5637细胞随着药物(顺铂3 μg/mL)作用时间增加,Cx43蛋白表达逐渐降低,差异有统计学意义,F=178.868,P＜0.001;而CleavedCaspase-3逐渐升高,差异有统计学意义,F=21.643,P＜0.001.顺铂(3μg/mL,4h)处理实验组和对照组后,CleavedCaspase-3蛋白相对表达量分别为0.740±0.092和0.373±0.091,差异统计学意义,t=6.394,P=0.001;BclL-2蛋白相对表达量分别为0.260±0.066和0.817±0.068,差异统计学意义,t=4.814,P=0.005 结论 沉默膀胱癌5637细胞中Cx43蛋白表达能提高顺铂的敏感性,可能与反常定位于细胞质中的Cx43参与线粒体介导的凋亡途径有关.     

Aberrant FHIT expression is linked to bladder carcinogenesis and apoptosis

The fragile histidine triad gene (FHIT) functions as tumor suppressor in many epithelial cell types. Although the exact mechanisms remain unclear, it is apparent that in its absence, cell cycle homeostasis is often perturbed resulting in the development of soft tissue tumors. Here, we investigated the role of FHIT expression in bladder carcinogenesis and progression using immunohistochemistry. Bladder carcinoma tissue and the 5637 cell line were also studied for FHIT expression by RT-PCR and Western blotting, respectively. FHIT was found to be expressed in carcinoma and adjacent normal tissues at both mRNA and protein levels, but the 17 kDa FHIT was lower in tumors (P<0.05), this being confirmed immunohistochemically. There was a negative correlation between FHIT expression and histological grade of bladder transitional cell carcinoma (P<0.05), but no clear relationship with clinical stage or relapse (P>0.05). Overexpression of FHIT could induce apoptosis in bladder carcinoma 5637 cells, which could be enhanced by adding adriamycin (ADR). These findings suggest important roles of FHIT in bladder cancer development and provide support for the feasibility of FHIT-based gene therapy.     

沉默IGF2R促进膀胱癌5637细胞增殖及肿瘤形成的机制探讨

目的:研究IGF2R在膀胱癌5637细胞中的功能。方法:应用Real-Time PCR和Western blot的方法检测膀胱癌组织及癌旁正常膀胱黏膜组织中IGF2R的表达水平。应用免疫荧光方法研究IGF2R在膀胱癌5637细胞中的定位情况。通过MTT法及体内裸鼠移植瘤的方法检测IGF2R对细胞增殖的影响。Western blot方法研究分子机制。结果:IGF2R在膀胱癌组织中呈现低表达(P<0.05)。体外及体内实验证明,沉默IGF2R表达可以促进膀胱癌5637细胞增殖(P<0.05),并且敲减IGF2R后我们观察到了磷酸化AKT水平的增高。结论:IGF2R在膀胱癌发生、发展过程中起到一个抑癌基因的作用。下调IGF2R可能通过激活AKT信号通路促进肿瘤细胞增殖。     

Coexistence of high levels of apoptotic signaling and inhibitor of apoptosis proteins in human tumor cells: implication for cancer specific therapy

It is well known that dysfunction of the apoptotic pathway confers apoptosis resistance and results in a low sensitivity of human cancer cells to therapeutic agents. A novel strategy to overcome the resistance is to target the apoptotic pathway directly. To identify molecular targets in the apoptotic pathway that are differentially regulated in cancer and normal cells, we have examined the levels of apoptotic effectors and inhibitors in human tumor and normal cell lines as well as in cancer and normal tissues. These include three pancreatic cancer lines (BXPC-3, MIA PaCa-2, and Panc-1), four breast cancer cell lines (MDA-MB-231, MDA-MB-435, MDA-MB-361, and MCF-7), and colon carcinoma line (SW620). Additionally, breast carcinoma tissue specimens were examined. Compared with normal human fibroblast and mammary epithelial cell lines, we detected high basal levels of caspase-3 and caspase-8 activities and active caspase-3 fragments in the tumor cell lines and cancer tissues in the absence of apoptotic stimuli. Furthermore, the tumor cells expressed high levels of survivin and XIAP, two members of the inhibitor of apoptosis (IAP) protein family. When the activity of these IAPs was blocked by expression of dominant-negative mutant survivin (survivinT34A) and XIAP-associated factor 1, respectively, apoptosis was induced in tumor but not normal cell lines. Moreover, down-regulation of both survivin and XIAP significantly enhanced tumor-cell apoptosis as compared with inhibition of either survivin or XIAP alone. These results suggest that up-regulated IAP expression counteracts the high basal caspase-3 activity observed in these tumor cells and that apoptosis in tumor cells but not normal cells can be induced by blocking IAP activity. Therefore, IAPs are important molecular targets for the development of cancer-specific therapeutic approaches.     

膀胱癌组织中DNMT蛋白表达水平与肿瘤增殖和凋亡程度的相关性

目的:探讨膀胱癌组织中DNMT蛋白表达水平与肿瘤增殖和凋亡程度的相关性。方法:收集膀胱癌组织标本80例和正常膀胱组织标本80例进行DNMT蛋白免疫组化分析。同时选择ABS-1膀胱癌细胞分为空白组、对照组、实验组。空白组加入常规培养基,对照组转染50nmol/L的空载体,实验组转染50nmol/L DNMT1 siRNA载体,然后进行肿瘤的增殖与凋亡分析。收集细胞进行DNMT蛋白表达检测。结果:DNMT1蛋白在膀胱癌组织中的阳性表达率为42.5%,在正常膀胱组织中的阳性表达率为2.5%,对比差异明显(P<0.05)。转染后DNMT1在空白组、对照组、实验组中的相对表达量分别为0.512±0.014、0.485±0.037和0.100±0.034,对比差异明显(P<0.05)。实验组增殖性明显低于空白组和对照组,而细胞凋亡率明显高于空白组和对照组,组间对比差异均有统计学意义(P<0.05)。结论:膀胱癌组织与膀胱癌细胞中存在DNMT1蛋白过表达,抑制DNMT1蛋白表达能有效抑制肿瘤细胞的增殖和促进凋亡。     

Survivin作为肿瘤标志物和治疗靶点的新进展

Survivin属于凋亡抑制蛋白(inhibi-tion of apoptosis protein,IAP)家族,能够抑制凋亡和调节分裂。近年来的研究显示,Sur-vivin在人类正常分化组织中很少表达却特异性表达在常见的肿瘤,它的表达上调可预示肿瘤患者生存期缩短和预后不良。作为肿瘤分子标志物,尿中高水平的Survivin表达能够辅助膀胱肿瘤的表达。应用新近的分子生物技术可以抑制肿瘤细胞中Survivin的表达,从而诱导肿瘤细胞的凋亡。     

膀胱癌组织GPR48表达及其临床意义

目的 G蛋白偶联受体48(G protein coupled receptor 48,GPR48)在人类细胞信号传导过程中起重要作用,并且与人类肿瘤的发生相关.本研究旨在探讨膀胱癌组织GPR48的表达水平,分析其与膀胱癌恶性程度的相关性.方法 选取2014 01-01 2016 05 20郑州大学附属郑州中心医院泌尿外科住院行手术切除治疗的膀胱癌组织标本60例和癌旁组织(距肿瘤组织＞2 cm)标本8例,BIU-87、5637和T24 3种膀胱癌细胞系作为研究对象,通过RT-PCR和蛋白质印迹法检测GPR48的表达量,收集60例患者性别、年龄、病理分级等临床病理资料,并检测其与GPR48表达的相关性.结果 GPR48在膀胱癌旁组织及癌旁组织中均有表达,膀胱癌组织和癌旁组织GPR48 mRNA表达量分别为0.080±0.048和0.041±0.032,差异有统计学意义,t=-3.047,P=0.005.膀胱癌组织和癌旁组织GPR48蛋白表达量分别为0.533±0.307和0.144±0.086,差异有统计学意义,t=-3.541,P=0.001.分析患者临床病理资料发现,GPR48的表达与患者性别无关,P＞0.05;而与患者年龄、病理分级和临床分期呈正相关,均P＜0.01.BIU-87、5637和T24细胞系中GPR48 mRNA的表达量分别为0.021±0.002、0.054±0.006和0.145±0.003,GPR48蛋白的表达量分别为0.718±0.076、0.905±0.121和1.205±0.08.T24细胞系GPR48 mRNA及蛋白表达量显著高于5637细胞系,均P＜0.05;5637细胞系GPR48 mRNA及蛋白表达量显著高于BIU-87细胞系,均P＜0.05.结论 GPR48表达与膀胱癌的恶性程度密切相关,有望成为一种潜在的生物标志物来预测膀胱癌的预后.     

赖氨酸甲基转移酶2D在膀胱癌中的表达水平及对细胞增殖和侵袭的影响

目的:检测赖氨酸甲基转移酶2D（KMT2D）在膀胱癌中的表达水平与临床病理特征的关系及KMT2D对膀胱癌细胞增殖和侵袭的影响。方法:免疫组化实验和qRT-PCR实验分别检测膀胱癌组织和癌旁组织中KMT2D蛋白和mRNA表达水平。根据免疫组化评分将患者分为KMT2D低表达组和KMT2D高表达组,比较两组患者的临床病理特征。蛋白免疫印迹实验检测SV-HUC-1、5637、T24和TCCSUP细胞中KMT2D蛋白表达水平。将miR-NC质粒和miR-KMT2D质粒转染至5637、T24和TCCSUP细胞中,蛋白免疫印记实验检测转染效率。MTT实验和Transwell实验检测细胞的增殖和侵袭情况。结果:膀胱癌组织中KMT2D蛋白免疫组化评分低于癌旁组织（t=17.371,P<0.001）。膀胱癌组织中KMT2D mRNA相对表达水平为（0.8±0.2）低于癌旁组织（2.6±0.4）,t=7.606,P=0.002;KMT2D低表达组与高表达组的年龄、肿瘤分化程度、淋巴结转移和浸润深度之间差异有统计学意义（P<0.05）;KMT2D蛋白在膀胱癌细胞5637、T24和TCCSUP中的表达水平分别为（0.38±0.03）、（0.42±0.03）、（0.36±0.05）均低于SV-HUC-1的表达水平（0.92±0.06）,q=21.131、19.584、21.647,均P<0.001;miR-KMT2D质粒转染至5637、T24和TCCSUP细胞后,KMT2D蛋白水平明显增加;细胞培养72 h时,5637-KMT2D组细胞OD值（0.73±0.05）低于5637-NC组（1.34±0.07）（t=8.241,P<0.001）,T24-KMT2D组细胞OD值（0.67±0.06）低于T24-NC组（1.12±0.06）（t=9.524,P<0.001）,TCCSUP-KMT2D组细胞OD值（0.62±0.07）低于TCCSUP-NC组（1.32±0.06）（t=8.941,P<0.001）;5637-KMT2D组细胞发生侵袭细胞数（34.5±4.7）低于5637-NC组（56.0±5.2）（t=3.524,P=0.008）,T24-KMT2D组细胞发生侵袭细胞数（36.4±4.5）低于T24-NC组（68.5±6.7）（t=7.302,P<0.001）,TCCSUP-KMT2D组细胞发生侵袭细胞数（30.1±3.4）低于TCCSUP-NC组（74.2±6.5）（t=8.206,P<0.001）。结论:KMT2D在膀胱癌组织中表达水平降低与患者的年龄、肿瘤分化程度、淋巴结转移和浸润深度有关,增加KMT2D的表达水平后可减弱细胞增殖和侵袭能力,KMT2D有望成为治疗膀胱癌的有效靶点。     

Expression of peroxisome proliferator-activated receptors (PPARs) in human urinary bladder carcinoma and growth inhibition by its agonists

Recent studies have demonstrated that peroxisome proliferator activator-receptors(PPAR)-gamma is expressed in various cancer tissues and its ligand induces growth arrest of these cancer cells through apoptosis. In our study, we investigated the expression of PPAR-alpha, beta and gamma in human bladder tumor (BT) and normal bladder (NB) tissues as well as the effects of PPAR-gamma ligands. Specimens were obtained from 170 patients with BT and 20 with NB. The expressions were investigated using RT-PCR and immunohistochemical methods. We also investigated the inhibitory effect of PPAR-gamma ligands on BT-derived cell line. Immunoreactive PPAR-alpha and -beta were significantly apparent in both BT and NB tissues. Although no marked expression of PPAR-gamma was observed in NB tissue, significant expression was found in BT tissue. The extent and intensity of immunoreactive PPAR-gamma polypeptides in BT cells were statistically much greater than those of NB cells. Correlation between PPAR-gamma expression and tissue type or progression of bladder cancer was observed; PPAR-gamma expression was higher in G3 of bladder cancer than in G1 and was higher in advanced than in early cancer. PPAR-gamma agonists, troglitazone and 15-deoxy-Delta(12, 14)-prostaglandin J(2) inhibited the growth of the BT cells. PPAR-gamma is expressed in bladder tumor, and results suggest that PPAR-gamma ligands may mediate potent antiproliferative effects against BT cells. Thus, PPAR-gamma has the ability to become a new target in treatment of bladder tumor.     

Elevated expression of inhibitor of apoptosis proteins in prostate cancer.

PURPOSE: Inhibitor of apoptosis (IAP) family proteins are suppressors of apoptosis that have been implicated in apoptosis resistance in some cancers. Their expression and relevance to the prognosis of prostate cancer were investigated. EXPERIMENTAL DESIGN: The expression of four members of the IAP family (cellular inhibitor of apoptosis protein 1, cellular inhibitor of apoptosis protein 2, X chromosome-linked IAP, and survivin) was examined by immunohistochemistry and immunoblotting in human prostate cancers and in prostate tissues from transgenic mice expressing SV40 large T antigen under control of a probasin promoter. RESULTS: Tumor-associated elevations in the levels of all four IAP family members were common in prostate cancers of both humans and mice, suggesting concomitant up-regulation of multiple IAP family proteins. Compared with normal prostatic epithelium, increased IAP expression was often evident even in prostatic intraepithelial neoplasia lesions (carcinoma in situ), suggesting that deregulation of IAP expression occurs early in the pathogenesis of prostate cancer. IAP expression did not correlate with Gleason grade or prostate-specific antigen levels. CONCLUSIONS: The findings demonstrate that tumor- associated elevations in the expression of several IAP family proteins occur as a frequent and early event in the etiology of prostate cancer.     

膀胱癌组织miR-451表达及其与E-cadherin和Vimentin表达相关性

目的 侵袭与转移是影响膀胱癌患者疗效及导致死亡的主要因素,其具体机制尚不明确;研究表明miR-451与多种肿瘤的发生、发展密切相关.本研究分析miR-451在膀胱癌中的表达及其临床意义,观察miR-451表达改变对上皮型钙黏附蛋白(E-cadherin)和间质型骨架蛋白(Vimentin)表达的影响.方法 收集2012-01-01-2015-12-31江西省人民医院收治的47例膀胱癌患者的癌组织及10例正常膀胱组织标本,运用实时荧光定量PCR (quantitative Real-time PCR,qPCR)检测miR-451的表达及运用免疫组织化学技术检测E-cadherin和Vimentin的表达,分析miR-451表达与膀胱癌临床病理因素,以及E-cadherin和Vimentin表达的关系.利用脂质体将miR-451拟似物(miR-451 mimics)转染入膀胱癌5637细胞,采用蛋白质印迹法检测E-cadherin和Vimentin的表达.结果 miR-451在膀胱癌组织中的表达为0.14±0.017,明显低于正常膀胱组织(1.0),其表达和膀胱癌临床分期及病理分级密切相关,P＜0.01;E-cadherin在膀胱癌组织较正常组织中低表达,分别为36.17％和84.50％;Vimentin在膀胱癌组织及正常膀胱黏膜层中表达不明显.癌组织中miR-451表达与E-cadherin表达正相关,r=0.67,P=0.023;miR-451过表达后,E-cadherin蛋白表达上调(P＜0.001),Vimentin蛋白表达无明显改变,P＞0.05.结论 MiR-451是膀胱癌潜在的调控子,可能是通过调控E-cadherin蛋白表达进而影响膀胱癌发生及进展.     

MicroRNA-30a在膀胱癌中的表达及作用

目的 探究微小RNA(miRNA)在人膀胱癌细胞系中的表达及其对人膀胱癌细胞增殖、凋亡及迁移侵袭能力的影响。方法 应用荧光实时定量PCR(qRT-PCR)法检测膀胱癌细胞系(5637和T24)和膀胱上皮永生化细胞(SV-HUC-1)中miRNA-30a的表达水平。通过对T24细胞转染miR-30a mimic和5637细胞转染miR-30a inhibitor上调或下调miR-30a的表达,并对其分别转染NC mimic和NC inhibitor作为对照。利用流式细胞技术、MTT法和Transwell法探究miR-30a的表达对膀胱癌细胞增殖、凋亡以及侵袭能力的影响。结果 在两种膀胱癌细胞系(5637和T24)中miRNA-30a的表达显著低于正常膀胱细胞系SV-HUC-1,并且在恶性度较高的T24膀胱癌细胞中其表达水平明显低于恶性度相对较低的5637细胞系。细胞转染72h后,miR-30a mimic组T24细胞OD值(0.83±0.09)明显低于NC mimic组(1.21±0.12)(P=0.003);miR-30a inhibitor组5637细胞OD值(1.28±0.14)高于NC inhibitor组(1.09±0.14)(P=0.019)。miR-30a mimic组T24细胞凋亡率(21.27±2.42)%明显高于NC mimic组(10.61±1.29)%;miR-30a inhibitor组5637细胞凋亡率(6.78±2.57)%明显低于NC mimic组(13.42±1.40)%,差异均具有统计学意义(P=0.0002,P=0.0014)。miR-30a mimic组穿膜细胞数(183.57±16.61)低于NC mimic组(465.80±9.20)(P＜0.0001);miR-30a inhibitor组(581.25±11.02)高于NC mimic组(397.13±7.57)(P＜0.0001)。结论 miR-30a表达的上调能够抑制膀胱癌细胞的增殖,促进细胞的凋亡,并降低膀胱癌细胞的迁移及侵袭的能力。膀胱癌细胞中miR-30a的低表达可能与膀胱癌的发生发展及转移有关。     

Down-Regulation of BAX Gene During Carcinogenesis and Acquisition of Resistance to 5-FU in Colorectal Cancer

Carcinogenesis and resistance to chemotherapy could be as results of expression variations in apoptosis regulating genes. Changes in the expression of apoptosis interfering genes may contribute to colorectal carcinogenesis and resistance to 5-Flourouracil (5-FU) during treatment schedule period. The present study aimed to evaluate the expression of pro-apoptotic and anti-apoptotic genes in colorectal cancer tumor tissues, normal adjacent tissues, and tumor colorectal cancer cell line during acquiring resistance to 5-FU in HT-29 based on Bolus treatment protocol. The normal and tumor tissues were obtained from hospital after surgery and total RNA was extracted for expression analysis. The HT-29 colorectal cancer cell line was cultured and exposed with 5-FU in three stages based on Bolus protocol. The MTT assay and Real Time PCR were carried out to determine the sensitivity to the drug and expression of desired genes, respectively. The obtained data showed that Proapoptotic genes, BAX and BID, were down-regulated in resistant derivate cells compared to wild type HT-29 cells. On the other hand Antiapoptotic genes, CIAP1 and XIAP, showed upregulation in resistant cells compared to wild type ones. Furthermore, BAX and FAS genes showed down-regulation in tumor samples in comparison to normal adjacent tissues. In conclusion, the results of our study suggest that BAX down-regulation could contribute as an important factor during both colorectal carcinogenesis and cell resistance to 5-FU.     

Expression and biological roles of SATB1 in human bladder cancer

Special AT-rich sequence-binding protein-1 (SATB1) has been recently reported to be overexpressed in various cancers and associate with the malignant behavior of cancer cells. However, the expression and potential roles of SATB1 in bladder cancer remains unclear. In the present study, SATB1 expression was analyzed in 85 archived bladder cancer specimens using immunohistochemistry and the correlations between SATB1 expression and clinicopathological parameters were evaluated. To further explore the biological functions of SATB1 in bladder cancer, siRNA knockdown was performed in 5637 and T24 bladder cancer cell lines. We then carried out CCK8 assay and examined cisplatin-induced apoptosis to address the roles of SATB1 in proliferation and apoptosis. We found that SATB1 was overexpressed in 33 of 85 (38.8 %) bladder cancer specimens. SATB1 overexpression associated with tumor grade (p?=?0.002) and tumor stage (p?=?0.027). SATB1 depletion in 5637 and T24 cells decreased cell proliferation while upregulating cisplatin-induced apoptosis. Further study demonstrated that SATB1 knockdown decreased cyclin D1 and cyclin E expression and upregulated caspase3 cleavage. In conclusion, SATB1 is overexpressed in bladder cancer and regulates malignant cell growth and apoptosis, which makes SATB1 a therapeutic target candidate for bladder cancer.     

靶向survivin基因siRNA对宫颈癌细胞放射敏感性影响的实验 

 目的 探讨survivin表达对宫颈癌放射敏感性的影响。 方法 RT-PCR和Western blot法分析survivin基因mRNA和蛋白的表达, 平板集落形成实验检测细胞 增殖,流式细胞术检测细胞凋亡率。 survivin siRNA转染HeLa细胞,比较survivin siRNA对细胞凋亡、增殖以及放射敏感性的 影响。 结果 在宫颈癌HeLa细胞中存在survivin的表达,survivin siRNA可诱导HeLa细胞凋亡并抑制增殖 。 结论 survivin siRNA可通过影响细胞凋亡和增殖来增加HeLa细胞的放射敏感性     

CTHRC1通过活化FAK-ERK1/2信号通路促进膀胱癌5637细胞迁移和侵袭

目的：探讨胶原三螺旋重复蛋白1(CTHRC1)在膀胱癌组织和细胞中的表达及其对膀胱癌5637细胞迁移和侵袭的影响及其机制。方法：利用TCGA和Arrayexpress数据库中膀胱癌基因表达数据,分析CTHRC1转录和翻译水平。收集2014年9月至2020年12月重庆医科大学附属第一医院手术切除的144例膀胱癌组织和25例全膀胱切除的癌旁组织标本,以及人膀胱癌细胞RT4、5637、T24、UMUC-3、TCCSUP和输尿管上皮永生化细胞SV-HUC-1。采用免疫组织化学染色法、qPCR法和WB法检测膀胱癌组织和细胞中CTHRC1的表达水平,通过Kaplan-Meier曲线分析CTHRC1表达对总生存期（OS）的影响。运用RNAi技术,敲降5637细胞CTHRC1表达后,通过细胞划痕实验和Transwell实验检测CTHRC1表达下调对5637细胞迁移和侵袭的影响。利用基因集富集分析（GSEA）预测CTHRC1相关的潜在信号通路,WB法检测敲降CTHRC1表达对FAK-ERK1/2通路相关蛋白表达的影响。结果：CTHRC1的转录和翻译水平在肌层浸润性膀胱癌（MIBC）组织和细胞中表达显著上...     

Antisense oligonucleotide targeting Livin induces apoptosis of human bladder cancer cell via a mechanism involving caspase 3

Background and Aim

in recent years, Livin, a new member of IAPs family, is found to be a key molecule in cancers. Researchers consider Livin may become a new target for tumor therapy; however, the role of it in bladder cancer is still unclear. The purpose of this article is to investigate Antisense Oligonucleotide (ASODN) of Livin on treating bladder cancer cell and underlying mechanisms.

Methods

Phosphorathioate modifying was used to synthesize antisense oligonucleotides targeting Livin, followed by transfection into human bladder cancer cell 5637. After transfection, Livin mRNA and protein level, cell proliferation and apoptosis changes, caspase3 level and its effect on human bladder cancer transplantable tumor in nude mice were measured.

Result

results showed Livin ASODN effectively inhibited Livin expression and tumor cell proliferation, and these effects probably through enhanced caspase3 activity and apoptosis of tumor cells. In nude mice transplantable tumor model, Livin expressions were inhibited meanwhile caspase3 expression was increased. Tumor growth slowed down and apoptosis was enhanced.

Conclusion

Our data suggest that Livin plays an important role in inhibiting apoptosis of bladder cancer cells. Livin ASODN may promote cell apoptosis, inhibit bladder cancer growth, and become one of the methods of gene therapy for bladder cancer.     

基于数据挖掘分析酪氨酸蛋白激酶受体TYRO3在膀胱癌中的表达及临床意义

目的:受体酪氨酸蛋白激酶亚家族TYRO3在多种肿瘤中高表达,但其在膀胱癌中未见系统研究,本文基于数据挖掘分析TYRO3在膀胱癌及正常组织中的表达情况,并进一步探讨TYRO3对膀胱癌预后的影响。方法:利用 Oncomine 数据库分析 TYRO3基因在膀胱癌组织中mRNA水平的变化。通过Fire Browse分析人体正常组织和其相应的肿瘤组织中TYRO3基因的表达差异。经MethHC分析膀胱癌和正常膀胱组织中TYRO3基因DNA启动子区甲基化水平的差异。利用OncoLnc做TYRO3基因的表达水平与膀胱癌患者生存期的相关性分析。在String-DB数据库中探索TYRO3 基因在细胞信号转导通路中的位置以及与其关系密切的上下游基因。结果:与正常膀胱组织相比,膀胱癌组织中TYRO3基因在mRNA水平呈高表达,TYRO3基因DNA启动子区甲基化水平低,TYRO3基因的表达水平与膀胱癌患者的总体生存时间呈负相关。GAS6、PHLPP1、PHLPP2、PIK3R1、PROS1、SOCS1、ACACA、ACACB、LRGUK、PI3、CCNH等基因与TYRO3有明显的相互作用。结论:大样本数据挖掘能迅速地获取浸润性膀胱癌组织中TYRO3表达的相关信息,为深入研究膀胱癌术后复发的监测和增加新的药物治疗靶点奠定基础。