Value of p16(INK4a) in the diagnosis of low-grade urothelial carcinoma of the urinary bladder in urinary cytology

BACKGROUND:

The sensitivity of urinary cytology for the diagnosis of urothelial carcinomas is low, particularly in low‐grade carcinomas. The UroVysion test is a fluorescent in situ hybridization multiprobe assay that increases the sensitivity of urinary cytology. However, this test is not widely available. P16^INK4a, a protein involved in cell cycle progression, is overexpressed in urothelial carcinoma. Immunocytochemical expression of p16^INK4a has been examined in biopsy samples from urothelial carcinomas, but few studies have addressed this protein in urine cytology.

METHODS:

The authors compared the results of p16^INK4a immunoreactivity in cytology and biopsy samples from 83 cases, including low‐grade urothelial carcinomas, reactive epithelial lesions, and negative cases.

RESULTS:

p16^INK4a assessment of in urine cytology samples showed a sensitivity of 66.7% and a specificity of 82.8% in the diagnosis of low‐grade urothelial carcinomas.

CONCLUSIONS:

On the basis of these results, the authors propose that immunocytochemical detection of p16^INK4a is a reliable tool in urine cytology, both for the diagnosis of low‐grade urothelial carcinomas and for follow‐up purposes. More retrospective and prospective studies are required to verify these results. Cancer (Cancer Cytopathol) 2012. © 2012 American Cancer Society.     

Urine and bladder washing cytology for detection of urothelial carcinoma: standard test with new possibilities

Background

Light microscopic evaluation of cell morphology in preparations from urine or bladder washing containing exfoliated cells is a standard and primary method for the detection of bladder cancer and also malignancy from other parts of the urinary tract. The cytopathologic examination is a valuable method to detect an early recurrence of malignancy or new primary carcinoma during the follow-up of patients after the treatment of bladder cancer.

Conclusions

Characteristic cellular and nuclear signs of malignancy indicate invasive or in situ urothelial carcinoma or high-grade papillary urothelial carcinoma. However, low sensitivity of the method reflects the unreliable cytopathologic diagnosis of low-grade urothelial neoplasms as cellular and nuclear signs of malignancy in these neoplasms are poorly manifested. Many different markers were developed to improve the diagnosis of bladder carcinoma on urinary samples. UroVysion™ test is among the newest and most promising tests. By the method of in situ hybridization one can detect specific cytogenetic changes of urothelial carcinoma.     

ADNP在膀胱尿路上皮癌组织中的表达及其临床意义

目的：探讨神经依赖性活性保护蛋白（ADNP）在膀胱尿路上皮癌中的表达情况及其临床意义。方法：收集中南大学湘雅医学院附属肿瘤医院2019 年6 月1 日至2019 年7 月15 日手术切除的膀胱癌及其配对的癌旁组织标本各28 例,采用qPCR检测20 例膀胱癌组织和癌旁组织的ADNP mRNA表达水平,WB检测其余8 对标本的ADNP蛋白表达水平。同时,回顾性分析我院2005 年1 月1 日至2007 年12 月31 日收治的膀胱尿路上皮癌患者221 例的临床病理资料,免疫组化染色方法检相应患者手术切除的石蜡标本中ADNP的表达情况,并收集同期因其他膀胱疾病而手术患者的非肿瘤膀胱组织切片用作对照。卡方检验分析ADNP表达与不同临床病理因素之间的相关性,Kaplan-Meier 法进行生存分析,Cox 比例风险回归模型对患者预后影响因素进行单因素及多因素分析。结果：膀胱尿路上皮癌组织中ADNP的转录和翻译水平均高于非肿瘤组织（均P<0.05）,且ADNP的表达量与膀胱癌的组织学分级、临床分期及患者存活状态有着相关性(P<0.05）。纳入的221 例患者随访中失访32 例,ADNP高表达较低表达的膀胱患者有着不良的预后（5 年OS：49.5% vs 78.6%,P<0.01;5 年PFS：40.0% vs 72.2% ,P<0.01;10 年OS：26.6% vs 58.6% ,P<0.01;10 年PFS：25.3% vs 47.9%,P<0.01）。Cox 单因素回归模型显示,ADNP表达量与膀胱癌的预后密切相关（P<0.05）;同时Cox 多因素回归也表明ADNP表达量（95% CI：1.300~2.905,P=0.001）是影响膀胱癌预后的独立危险因素。结论：ADNP在膀胱癌组织中的表达水平比非肿瘤的膀胱组织有显著的升高,组织学分级及临床分期与ADNP的表达水平有相关性,ADNP低表达的膀胱癌患者预后相对较好,ADNP有望成为膀胱癌的特异性治疗候选靶点。     

miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭

背景与目的：生物信息学分析提示GATA6是miR-203a-3p的潜在靶基因,明确miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭。方法：采用Lipofectamine ^TM RNAiMAX对培养的KYSE-70和KYSE-180细胞瞬时转染。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）检测miR-203a-3p和GATA6的表达水平。采用蛋白质印迹法（Western blot）检测GATA6蛋白的水平。质粒联合转染后检测相对萤光素酶活性。对食管鳞癌患者标本进行恶性肿瘤与异型增生组织miR-203a-3p和GATA6的表达检测。结果：RTFQ-PCR及Western blot检测结果显示,与对照组比较,GATA6基因和蛋白的表达在miR-203a-3p转染组中降低,在miR-203a-3p inhibitor组却升高,差异均有统计学意义（P<0.05）。与对照组比较,miR-203a-3p转染组KYSE-70细胞增殖能力下降,miR-203a-3p inhibitor组中却升高,差异均有统计学意义（P<0.05）。在KYSE-180中虽然差异无统计学意义,但其趋势却与KYSE-70一致。与对照组比较,在KYSE-70和KYSE-180细胞系中,侵袭细胞数值/视野在miR-203a-3p转染组中均明显下降（P<0.01）,在miR-203a-3p inhibitor组中却明显升高（P<0.05）。与miR-203a-3p掠夺型+GATA6野生型组和miR-203a-3p野生型+GATA6突变型组比较,相对萤光素酶活性在miR-203a-3p野生型+GATA6野生型组中降低,差异有统计学意义（P<0.05）。与异型增生组织相比较,100%（10/10）食管鳞癌患者miR-203a-3p在恶性肿瘤组织的表达下调,而GATA6表达水平上调。结论：miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭能力。     

Upregulation of TRAG3 gene in urothelial carcinoma of the bladder

Conventional chemotherapy is commonly used for advanced stages of bladder cancer with modest success and high morbidity. Identifying markers of resistance will allow clinicians to tailor treatment to a specific patient population. T24‐tumorigenic cell line was grown orthotopically in nude mice and monitored using bioluminescence imaging and microcomputed tomography until they developed metastases. Stable sublines were then developed from primary bladder (T24‐P), lung (T24‐L) and bone (T24‐B) tissues. Chromosomal analysis and DNA microarray were used to characterize these sublines. Real‐time quantitative polymerase chain reaction and immunohistochemistry were used for validation. Epigenetic modifiers were used to study gene regulation. The cell viability was quantified with MTT assay. Chromosomal analysis revealed multiple alterations in metastatic cell lines compared to T24‐P. DNA microarray analysis showed that taxol resistance‐associated gene (TRAG) 3 was the most upregulated gene. From real‐time quantitative polymerase chain reaction and immunohistochemistry, TRAG3 was significantly higher in T24‐L and T24‐B than T24‐P. TRAG3 gene expression is likely controlled by DNA methylation but not histone acetylation. Interestingly, T24‐B and T24‐L cells were more resistant than T24‐P to treatment with antimicrotubule agents such as docetaxel, paclitaxel and vinblastine. TRAG3 mRNA expression was higher in 20% of patients with ≤pT2 (n = 10) and 60% of patients with ≥pT3 (n = 20) compared to normal adjacent tissue (p = 0.05). In addition, the median TRAG3 expression was 6.7‐fold higher in ≥pT3 tumors compared to ≤pT2 tumors. Knowing the status of TRAG3 expression could help clinicians tailor treatment to a particular patient population that could benefit from treatment, while allocating patients with resistant tumors to new experimental therapies.