Study of wavy laminar growth of human urinary bladder cancer cell line in vitro

Objective: To observe the ordered growth behavior of human urinary bladder cancer cell line (BIU) under culture in vitro. Methods: The suspension of BIU cells was spread locally in a culture container. When the cells grew a-long the wall to form a cellular colony, macroscopic and microscopic observations complemented with measurements of the parameters including expanding diameter, expanding rate, cell shape. average cell density, average cell size. dehydrogenase activity and sensitivity to pH were conducted dynamically. Results: During cell culture, obvious laminar characteristics appeared in localized growing BIU cell colonies and there was difference between the cells of different zones in shape, size, density, dehydrogenase activity and sensitivity to pH. Conclusion: Space closing and bio-dissipation result in self-organization of BIU cells with ordered growth behavior. The present experiment offers a simple, controllable model for the study of wavy growth of human cells.

Effect of Photodynamic Therapy with BPD-MA on the Proliferation and Apoptosis of Human Bladder Cancer Cells

OBJECTIVE To explore the effect of photodynamic therapy with benzoporphyrin derivative monoacid ring A （BPD-MA） on the proliferation and apoptosis of human bladder cancer cells. METHODS Photosensitization of BPD-MA was activated with a red light laser （632.8 nm） delivered at 10 mw/cm^2 to give a total dose of 2.4 J/cm^2. Cellular proliferative activity was measured using the 3-（4,5.- dimethylethiazil-2-yl）-2,5-Diph3-eyl tetrazolium bromide （MFi-） assay and 3H-thymidine incorporation. Cell apoptosis was determined with flow cytometry analysis and the terminal deoxyuridine nicked-labeling （TUNEL） assay. RESULTS At 24 h post photodynamic treatment, photodynamic therapy significantly decreased cellular proliferative activity. The rate of apoptosis in BIU-87 cells 8 h after photodynamic treatment significantly increased up to 26.11± 2.59% as analyzed with flow cytometry. In situ labeling of DNA cleavage products with the terminal deoxyuridine nicked-labeling （TUNEL） assay reinforced these observations, BPD-MA-mediated photosensitization increased the number of TUNEL-positive cells compared to the controls. However, laser irradiation alone, BPD-MA alone and sham radiation did not affect cellular proliferative activity or apoptosis of the human bladder cancer BIU-87 cells. CONCLUSION Photodynamic therapy with BPD-MA significantly decreases cellular proliferative activity and enhances apoptosis. Therapy using this method might be a promising approach to treat patients with bladder cancer.

Stem cell applications for pathologies of the urinary bladder

New stem cell based therapies are undergoing intense research and are widely investigated in clinical fields including the urinary system. The urinary bladder performs critical complex functions that rely on its highly coordinated anatomical composition and multiplex of regulatory mechanisms. Bladder pathologies resulting in severe dysfunction are common clinical encounter and often cause significant impairment of patient's quality of life. Current surgical and medical interventions to correct urinary dysfunction or to replace an absent or defective bladder are sub-optimal and are associated with notable complications. As a result, stem cell based therapies for the urinary bladder are hoped to offer new venues that could make up for limitations of existing therapies. In this article, we review research efforts that describe the use of different types of stem cells in bladder reconstruction, urinary incontinence and retention disorders. In particular, stress urinary incontinence has been a popular target for stem cell based therapies in reported clinical trials. Furthermore, we discuss the relevance of the cancer stem cell hypothesis to the development of bladder cancer. A key subject that should not be overlooked is the safety and quality of stem cell based therapies introduced to human subjects either in a research or a clinical context.

Effects of Combined siRNA-TR and-TERT on Telomerase Activity and Growth of Bladder Transitional Cell Cancer BIU-87 Cells

The effects of combined RNA interference(RNAi) of human telomerase RNA(hTR) and human telomerase reverse transcriptase(hTERT) genes on telomerase activity in a bladder cancer cell line(BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer(BTCC).Three TR-specific double-stranded small interfering RNAs(siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA.The phTR-siRNA,phTERT-siRNA,and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells.The expression of hTR and hTERT mRNA was detected by quantitative fluorescent reverse transcription-polymerase chain reaction,and a telomeric repeat amplification protocol was applied to detect telomerase activity.Growth inhibition of BIU-87 cells was measured by MTT assay.Gene chip analysis was performed to evaluate the effects of the combined RNAi of hTR+hTERT genes on telomerase activity and growth of BIU-87 cells in vitro.The results showed that the expression of hTERT and hTR mRNA was inhibited by pRNAT-hTERT-Ⅲ,pRNAT-hTR-Ⅲ,and pRNAT-hTR-Ⅲ+hTERT-Ⅲ in BIU-87 cells.The inhibition efficiency of pRNAT-hTERT-Ⅲ,pRNAT-hTR-Ⅲ,pRNAT-hTERT-Ⅲ+pRNAT-hTR-Ⅲ was 67% for TERT mRNA,41% for TR mRNA,57% for TR mRNA and 70% for TERT mRNA in BIU-87 cells respectively.The growth of BIU-87 cells was inhibited and telomerase activity was considerably decreased,especially in the cells treated with combined RNAi-hTR and-hTERT.Gene chip analysis revealed that 21 genes were down-regulated(ATM,BAX,BCL2,BCL2L1,BIRC5,CD44,CTNNB1,E2F1,JUN,MCAM,MTA1,MYC,NFKB1,NFKBIA,NME4,PNN,PNN,SERPINE1,THBS1,TNFRSF1A,and UCC1).The results indicated that hTR-siRNA and hTERT-siRNA,especially their combination,siRNA hTR+hTERT,specifically and effectively suppressed the expression of both hTR and hTERT mRNA and telomerase activity.Molecular biological mechanism by which combined siRNA-TR and-TERT inhibited telomerase activity and growth of BIU-87 cells in vitro may involve the down-regulation of the 21 genes.

Understanding the role of transmembrane 9 superfamily member 1 in bladder cancer pathogenesis

In this editorial we comment on the article by Wei et al,published in the recent issue of the World Journal of Clinical Oncology.The authors investigated the role of Transmembrane 9 superfamily member 1(TM9SF1)protein in bladder cancer(BC)carcinogenesis.Lentiviral vectors were used to achieve silencing or overexpression of TM9SF1 gene in three BC cell lines.These cell lines were then subject to cell counting kit 8,wound-healing assay,transwell assay,and flow cytometry.Proliferation,migration,and invasion of BC cells were increased in cell lines subjected to TM9SF1 overexpression.TM9SF1 silencing inhibited proliferation,migration and invasion of BC cells.The authors conclude that TM9SF1 may be an oncogene in bladder cancer pathogenesis.

Apoptosis Induced by Photodynamic Therapy with Benzoporphyrin Derivative Monoacid Ring A and Exploration of its Potential Mechanism in Bladder Cancer Cells

OBJECTIVE To investigate apoptosis induced by photodynamic therapy with benzoporphyrin derivative monoacid ring A （BPD-MA） and explore its potential mechanism in human bladder cancer cells. METHODS Photosensitization of BPD-MA was activated with a red light Laser （632.8nm） delivered at 10 mW/cm^2 to give a total dose of 2.4 J/cm^2. Cellular apoptosis was measured with flow cytometry analysis and an insitu terminal deoxyuridine nick end-labeling （TUNEL） assay. Changes in mitochondrial membrane potential （△φm） were monitored by a flow cy-tometric method with Rhodamine 123 staining and the expression of bcl- 2 in BIU-87 cells was detected with immunocytochemical staining. RESULTS At 8 h following photodynamic treatment, the degree of apoptosis was significantly increased when analyzed with flow cytometry and TUNEL assay. Treatment of the BIU-87 cells by PDT with BPD-MA resulted in the collapse of the △φm and a decrease of bcl-2 expression. CONCLUSION BPD-MA-mediated PDT can effectively induce apoptosis in BIU-87 cells. The mechanism probably is through a mitochondrial-initiated pathway.

Tissue-engineered conduit using bladder acellular matrix and bladder epithelial cells for urinary diversion in rabbits

Background For muscle invasive bladder cancer, radical cystectomy is the most effective treatment now and urinary diversion is often necessary. The use of intestinal tissue for urinary diversion is frequently associated with complications. In this study, we aimed to make a tissue-engineered conduit （TEC） using bladder epithelial cells and bladder acellular matrix （BAM） for urinary diversion in rabbits. Methods Bladder epithelial cells of rabbit were cultivated and expanded in vitro, then seeded on BAM, and cultured for 7 days. Then cell-seeded graft was used to make TEC. In the experimental group, most of bladder of the rabbit was removed while bladder trigone was retained. The proximal end of TEC was anastomosed with bladder trigone and the distal end was anastomosed with the abdominal stoma. In the control group, TEC was made using unseeded BAM. Haematoxylin and eosin staining was conducted, respectively, at 1, 2, 4, and 8 weeks postoperatively. Immunohistochemistry was performed 8 weeks postoperatively. Intravenous urography, retrograde pyelography, and cystoscopy of TEC were made at 12 weeks postoperatively. Results All animals were alive in the experimental group. Haematoxylin and eosin staining showed epithelial coverage in TEC. Immunohistochemistry showed anti-cytokeratin AEI/AE3 antibody and anti-ZO1 antibody positive, confirming there were mature and functional epithelial cells on the lumen of TEC. Retrograde pyelography and intravenous urography showed that TEC developed well and that there was no obstruction. In the control group, four rabbits were dead within 2 weeks and scar formation, atresia, and severe hydronephrosis were found. Conclusions We successfully made TEC using BAM and bladder epithelial cells for urinary diversion in rabbits. The lumen of this new TEC covered mature epithelial cells and could prevent urinary extravasation.

TGF-β对膀胱癌组织中巨噬细胞来源肿瘤相关成纤维细胞的影响及其机制

目的探讨TGF-β对膀胱癌组织中巨噬细胞来源肿瘤相关成纤维细胞(CAF)的影响及其机制。方法采用Kaplan-Meier法分析膀胱癌组织中α-SMA^(+)CD68^(+)CAF表达水平与患者的总生存率(OS)的关系;采用免疫荧光技术检测α-SMA^(+)CD68^(+)CAF在膀胱癌组织中的浸润情况;采用Western blot实验检测TGF-β对α-SMA^(+)CD68^(+)CAF体外诱导作用。同时构建膀胱癌小鼠模型,采用免疫荧光技术和免疫组化法检测TGF-β对膀胱癌小鼠体内α-SMA^(+)CD68^(+)CAF的诱导作用及对CD8^(+)T细胞浸润的影响。结果膀胱癌组织中α-SMA与CD68的表达呈正相关,且α-SMA^(+)CD68^(+)CAF高表达的患者预后更差(χ^(2)=9.05,P<0.05)。免疫荧光技术检测结果显示,膀胱癌组织中存在α-SMA^(+)CD68^(+)CAF。Western blot实验检测结果显示,TGF-β可显著促进α-SMA^(+)CD68^(+)CAF的生成。膀胱癌小鼠模型体内实验显示,TGF-β可促进膀胱癌组织中α-SMA^(+)CD68^(+)CAF的生成,从而抑制CD8^(+)T细胞的浸润。结论TGF-β可显著促进膀胱癌组织中巨噬细胞来源CAF的生成,从而抑制CD8^(+)T细胞的浸润。

罗哌卡因抑制膀胱癌J82细胞增殖、迁移及侵袭的分子机制研究

目的探讨罗哌卡因对人膀胱癌J82细胞增殖、迁移和侵袭的影响及对c-Jun氨基末端激酶(JNK)信号通路的调控作用。方法体外培养人膀胱癌J82细胞,该研究分为预实验和正式实验两个部分。预实验分组:对照组(不进行干预),低、中、高浓度罗哌卡因组(200、400、800μmol/L罗哌卡因);正式实验分组:对照组、罗哌卡因组(有显著作用浓度的罗哌卡因)、阳性药物组(100 nmol/L紫杉醇)、抑制剂组(罗哌卡因+20μmol/L JNK信号通路抑制剂SP600125)和激活剂组(罗哌卡因+1μg/mL JNK信号通路激活剂茴香霉素),干预24 h。分别采用细胞计数试剂盒-8(CCK-8)法、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)、划痕法、Transwell小室法、蛋白免疫印迹法检测细胞活力、增殖率、迁移率、侵袭细胞数、上皮间质转化(EMT)及JNK通路相关蛋白水平。结果预实验中,与对照组比较,高浓度罗哌卡因组细胞活力明显降低(P<0.05),以高浓度罗哌卡因(800μmol/L)进行正式实验。正式实验中,罗哌卡因组和阳性药物组细胞增殖率、迁移率、侵袭细胞数,以及N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、纤维粘连蛋白(FN)和磷酸化(p)-JNK表达水平较对照组降低(P<0.05),E-钙黏蛋白(E-cadherin)表达水平较对照组升高(P<0.05)。与罗哌卡因组相比,抑制剂组细胞增殖率、迁移率、侵袭细胞数,以及N-cadherin、Vimentin、FN和p-JNK表达水平降低(P<0.05),E-cadherin表达水平升高(P<0.05),而激活剂组上述指标趋势则正好相反(P<0.05)。结论罗哌卡因可能通过阻断JNK信号通路活化抑制人膀胱癌J82细胞增殖、迁移、侵袭和EMT进程。

PEG-rhG-CSF预防小细胞肺癌化疗后中性粒细胞减少症的效果分析

目的:探讨聚乙二醇化重组人粒细胞刺激因子(PEG-rhG-CSF)预防小细胞肺癌化疗后中性粒细胞减少症的效果。方法:选取2020年1月—2023年3月北京市朝阳区桓兴肿瘤医院收治的小细胞肺癌化疗患者200例,按照信封法分为两组。对照组(n=98)化疗后应用重组人粒细胞刺激因子(rhGCSF),观察组(n=102)化疗后应用PEG-rhG-CSF。比较两组中性粒细胞减少症发生率、免疫功能、血清肿瘤标志物水平及不良反应发生情况。结果:观察组中性粒细胞减少症发生率低于对照组,差异有统计学意义(P<0.05)。治疗后,观察组CD3^(+)、CD4^(+)水平均高于对照组,CD8^(+)及细胞角质蛋白19片段(Cyfra21-1)、胃泌素释放肽前体(Pro-GRP)、神经元特异性烯醇化酶(NSE)水平均低于对照组,差异均有统计学意义(P<0.05)。两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论:小细胞肺癌化疗后应用PEG-rhG-CSF能有效预防中性粒细胞减少症的发生,改善机体免疫功能,抑制血清肿瘤标志物水平,且安全性良好。

经完全性手术切除的Ⅱ~ⅢA期EGFR野生型NSCLC术后辅助化疗中预防应用PEG-rhG-CSF的临床获益分析

目的探究经完全性手术切除的Ⅱ~ⅢA期表皮生长因子受体(EGFR)野生型非小细胞肺癌(NSCLC)术后辅助化疗中预防应用聚乙二醇化重组人粒细胞集落刺激因子(PEG-rhG-CSF)的临床获益。方法前瞻性选取2019年2月至2021年2月广东医科大学附属医院经完全性手术切除的Ⅱ~ⅢA期EGFR野生型NSCLC术后辅助化疗患者90例作为研究对象,根据随机数字表法将患者分为观察组与对照组,每组各45例。对照组预防性应用重组人粒细胞(rhG-CSF),观察组预防性应用PEG-rhG-CSF。对患者随访3个月,比较两组治疗前及治疗1、3、5、10 d后的白细胞计数、中性粒细胞计数水平及白细胞减少症、中性粒细胞减少症发生情况,并记录患者随访期内化疗后的不良反应情况。结果两组患者在治疗前及治疗1、3、5 d后的白细胞计数水平比较,差异均无统计学意义(P>0.05);治疗10 d后,观察组患者的白细胞计数水平为(5.65±1.11)×10^(9)/L,高于对照组[(4.66±1.08)×10^(9)/L],差异有统计学意义(P<0.05)。两组患者在治疗前及治疗1、3、5 d后的中性粒细胞计数水平比较,差异均无统计学意义(P>0.05);治疗10 d后,观察组患者的中性粒细胞计数水平为(3.61±1.51)×10^(9)/L,高于对照组[(2.65±1.46)×10^(9)/L],差异有统计学意义(P<0.05)。观察组患者2、3级白细胞减少症和中性粒细胞减少症发生率分别为15.56%、11.11%,均低于对照组(35.56%、42.22%),差异均有统计学意义(P<0.05)。两组患者在骨骼肌疼痛、疲劳、感染、发热、胃肠反应等方面比较,差异均无统计学意义(P>0.05)。结论对经完全性手术切除后Ⅱ~ⅢA期EGFR野生型NSCLC患者,术后辅助化疗中预防应用PEG-rhG-CSF可以提高白细胞计数和中性粒细胞计数水平,并减少白细胞减少症和中性粒细胞减少症的发生,有助于改善患者的免疫功能和预后。

三个膀胱癌细胞系侵袭相关分子的研究

目的 探讨膀胱癌细胞侵袭能力不同的机制。方法 取来源于同一膀胱癌组织的三个细胞系 ,检测其软琼脂克隆形成率及基底膜黏附能力。采用荧光偏振法测定细胞膜脂流动性 ,免疫组化法和流式细胞术分析E cadherin、基质金属蛋白酶 2 (matrixmetalloproteinases 2 ,MMP 2 )和CD44的表达。 结果 BLX和BLZ细胞的软琼脂克隆形成率高于BLS细胞 ,同时基底膜黏附能力均大于BLS细胞 ,BLX和BLZ细胞的膜脂流动性小于BLS细胞 ,BLX、BLZ和BLS细胞在MMP 2表达上无差异 ,BLS和BLZ的E cadherin表达强于BLX细胞。流式细胞仪分析BLS细胞CD44阳性率高于BLX和BLZ细胞。

重组人粒细胞集落刺激因子(rhG-CSF)对小细胞肺癌化疗后骨髓抑制的有效性和安全性分析

目的评价重组人粒细胞集落刺激因子(rhG-CSF,造血生长因子)对小细胞肺癌化疗所致骨髓抑制的疗效。方法对解放军总医院2002年至2006年来222例小细胞肺癌患者病例进行回顾性调查,采集其中62例化疗后使用重组人粒细胞集落刺激因子进行治疗的病例,并对信息进行统计学分析。结果重组人粒细胞集落刺激因子可以有效缓解小细胞肺癌患者化疗后的骨髓抑制反应,使白细胞和中性粒细胞数目明显升高,平均恢复时间3天,有效率为95.2%。结论rhG-CSF对治疗小细胞肺癌化疗所致的骨髓抑制作用好,不良反应少;但对不同患者的不同情况,应调整剂量和用法。

人膀胱癌细胞波状层次生长的研究

目的 观察人膀胱癌细胞 (BIU)在体外培养条件下的生长有序行为。方法 将BIU细胞悬液进行局限区域接种 ,待其形成贴壁生长的群落后进行宏观和显微动态观察及参数测量 (扩展直径、扩展速率、细胞形态、细胞平均密度、细胞平均大小、脱氢酶活性、对pH的敏感性等 )。结果 体外细胞培养条件下 ,局限区域生长的BIU细胞群落能出现明显的层次特性 ,不同区域细胞的形态、大小、密度、脱氢酶活性以及对pH的敏感性均存在差异。结论 空间封闭和生物耗散导致细胞自组织 ,其结果表现为生长有序行为。

VX-680诱导人膀胱癌T24细胞凋亡及其对Bcl-2表达的影响

目的:探讨Aurora A激酶抑制剂VX-680对人膀胱癌T24细胞凋亡的诱导作用及对Bcl-2 mRNA和蛋白表达的影响及意义。方法:体外培养T24细胞,分别给予不同浓度及作用时间的VX-680,Annexin V/PI检测细胞凋亡;Hoechst染色法检测细胞凋亡形态学改变;RT-PCR和Western blot检测细胞Bcl-2 mRNA和蛋白的表达。结果:Annexin V/PI显示,凋亡率分别随着浓度增高和时间增长而增加,呈浓度及时间依赖性;Hoechst染色发现细胞在处理72 h后,其凋亡增多程度及Bcl-2 mRNA和蛋白表达下调水平均随浓度增高而增加,呈浓度依赖性。结论:Aurora激酶抑制剂VX-680可能通过下调Bcl-2表达显著诱导人膀胱癌T24细胞凋亡,且具有浓度依赖性。

木犀草素联合卡介苗治疗膀胱癌的协同效应及其机制

目的:研究木犀草素(Luteolin,Lu)联合卡介苗(bacillus calmette-guerin,BCG)抑制膀胱癌BIU-87细胞增殖的作用及其机制。方法:体外培养膀胱癌BIU-87细胞株,分别用不同浓度的Lu(20,40,60,80,100,160μmol/L)单独及联合BCG(100,200 mg/L)处理BIU-87细胞,作用6,12,24,48 h。采用Hoechst 33258核荧光染色观察细胞形态学变化,MTT法检测Lu和BCG对BIU-87细胞增殖的抑制作用和半数抑制率IC50,流式细胞术分析肿瘤细胞的凋亡及细胞周期,比色法测定caspase-3蛋白活性,Western印迹分析磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinases,P-JNK)的表达。结果:Hoechst 33258核荧光染色提示Lu可诱导细胞凋亡,并增强BCG诱导的细胞凋亡。MTT法检测显示Lu及BCG对BIU-87细胞增殖均有明显抑制作用,且呈浓度和时间依赖性;Lu与BCG联合作用对BIU-87细胞的增殖抑制具有显著的协同效应(P<0.05)。流式细胞术提示Lu及BCG均诱导细胞周期阻滞及细胞凋亡,并具有明显的协同增敏效应(P<0.05)。Lu及BCG均可上调caspase-3活性及P-JNK表达水平,二者联合作用明显增强(P<0.05)。结论:Lu与BCG均可抑制BIU-87细胞增殖,诱导细胞凋亡,并呈浓度依赖性,二者具有明显的协同增敏作用;Caspase-3蛋白活化及JNK的激活是其可能的分子机制,Lu可作为BCG免疫治疗的有效增敏剂用于对膀胱肿瘤的治疗。

苦参碱对人膀胱癌BIU-87细胞增殖的抑制作用及其机制研究

目的：研究苦参碱对人膀胱癌BIU-87细胞增殖的抑制作用及其机制。方法：以0（阴性对照）、0.5、1.0、2.0、4.0 mg/ml苦参碱分别作用于人膀胱癌BIU-87细胞24、48、72 h后,MTT法检测细胞活性并计算细胞增殖抑制率;以0（阴性对照）、0.5、1.0、2.0、4.0 mg/ml苦参碱作用于细胞48 h后,流式细胞术检测细胞周期和凋亡率,Western blot法检测细胞中生存素、天冬氨酸半胱氨酸蛋白酶（Caspase）-3及Caspase-7蛋白的表达。结果：与阴性对照比较,1.0-4.0 mg/ml苦参碱作用24、48、72 h后均对BIU-87细胞增殖有显著的抑制作用（P〈0.05或P〈0.01）,增殖抑制率呈浓度和时间依赖性升高;作用48 h后,G0/G1期细胞比例升高、S期及G2/M期细胞比例降低、凋亡率升高（P〈0.05或P〈0.01）。0.5-4.0 mg/ml苦参碱作用48 h后,细胞中生存素蛋白表达减弱、Caspase-3及Caspase-7蛋白表达增强（P〈0.05或P〈0.01）。结论：苦参碱可抑制人膀胱癌BIU-87细胞的增殖、阻滞细胞周期、诱导细胞凋亡,其机制可能与调控细胞中生存素及Caspase-3、Caspase-7的表达有关。

集落刺激因子1及其受体的单抗对人肝癌细胞在裸鼠体内生长的抑制作用

探讨集落刺激因子┐１及其受体在人原发性肝癌中的过量表达及其意义。方法抗ＣＳＦ１Ｒ和抗ＣＳＦ１单克隆抗体对裸鼠移植性人肝癌７７２１细胞的生物学作用。结果两个单抗分别以７４．８８％和７７．９３％的体积抑瘤率在裸鼠体内抑制７７２１细胞的生长。结论ＣＳＦ１Ｒ和ＣＳＦ１可能是一新发现的肝癌自泌和旁泌系统。

RNA干扰MEX3A基因对膀胱癌细胞增殖和凋亡的影响

目的探讨通过慢病毒介导的RNA干扰技术抑制MEX3A基因表达对膀胱癌细胞增殖和凋亡的影响。方法应用GV115载体构建针对MEX3A基因的shRNA慢病毒载体,转染包装细胞293T产生慢病毒颗粒,收集并滴度测定后转染5637膀胱癌细胞,实验组转染MEX3A-shRNA慢病毒,对照组转染阴性对照慢病毒;实时定量PCR(Real-time PCR)检测MEX3A基因敲减效率;慢病毒转染3 d后,使用Celigo连续细胞计数5d检测细胞生长;慢病毒转染5d后,进行膜联蛋白V(Annexin V)染色并用流式细胞术检测细胞凋亡。结果成功构建MEX3AshRNA慢病毒载体以及稳定抑制MEX3A表达的细胞株,MEX3A基因敲减效率达到74%;Celigo细胞计数检测显示,与对照组相比,实验组5637细胞的增殖速率受到显著抑制;流式细胞术检测显示,实验组发生凋亡的5637细胞显著增加。结论 MEX3A基因促进膀胱癌细胞的增殖,抑制膀胱癌细胞的凋亡。

自动尿液细胞DNA定量分析对泌尿系统炎症与膀胱癌的鉴别诊断价值

目的:评估尿液细胞DNA定量分析在泌尿系统肿瘤诊断中的应用价值。方法:采集92例泌尿系统炎症和39例疑似膀胱癌患者的尿液分别进行DNA定量测定、常规细胞学检验和尿液液基细胞学检查,以病理诊断为金标准,评估3种方法对泌尿系统炎症与膀胱癌的鉴别诊断价值,分析尿液细胞DNA定量与肿瘤分型的关联。结果:尿液DNA倍体分析对膀胱癌诊断的敏感性和特异性分别为88.89%和97.09%,诊断符合率为94.66%。传统尿液细胞学检验的敏感性和特异性分别为51.85%和100%,准确性为90.07%。尿液液基细胞学检查对膀胱癌诊断的敏感性和特异性分别为81.48%和99.04%,诊断符合率为95.41%。尿液DNA倍体分析结果>5C细胞的个数与尿路上皮癌分型存在关联,高级别尿路上皮癌组异倍体(>5C)细胞数量较低级别组显著增高。结论:尿液DNA定量分析能够较好地鉴别泌尿系统炎症与膀胱癌,较传统细胞学检验有更好的提示作用。