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Objective:To investigate the correlation of intratumoral fibrosis with the prognosis of clear cell renal cell carcinoma (ccRCC).Methods:The correlation of the transcriptional expression of the primary collagen with the prognosis in ccRCC was evaluated using the Cancer Genome Atlas (TCGA) database, including 530 ccRCC patients with complete information. Of them, 344 cases were male, 186 cases were female. The age of 264 cases was ≤ 60 years, and the age of 266 cases was > 60 years. The pathology grade of 241 patients was G 1-2 grade, and the pathology of 281 cases were G 3-4 grade, 8 cases were undetermined grade. There were 322 cases with AJCC stage Ⅰ-Ⅱ and 205 cases with AJCC stage Ⅲ-Ⅳ, and 3 cases with undetermined stage. There were 420 cases in M 0 and 78 cases in M 1, and 32 cases without distant metastases information. Furthermore, the paraffin sections of 158 non-cystic ccRCC patients confirmed by pathology from November 2005 to November 2017 were further used to evaluate the level of collagen of ccRCC and the status of the pseudocapsule by the Masson staining, Sirius red staining and multicolor immunofluorescence staining of collagen Ⅰ and collagen Ⅲ. Of them, 112 cases were male, 46 cases were female. There were 100 cases with age ≤ 60 years, and 58 cases with age > 60 years. The pathology grade of 111 cases were G 1-2, and the pathology grade of 47 cases were G 3-4. There were 144 cases with AJCC stage Ⅰ-Ⅱ, 14 cases with AJCC stage Ⅲ-Ⅳ. Kaplan-Meier survival curve were used to analyze the relationship between tumor collagen parameters and the overall survival prognosis of patients with ccRCC. Results:The transcriptome results of the TCGA database indicated that the expression level of COL1A1 in ccRCC tissues was significantly higher than that in adjacent normal tissues ( P<0.001). The high expression of collagen suggested a worse overall survival prognosis ( HR=1.165, P=0.002). In addition, the high ratio of COL1A1/COL3A1 indicated a worse overall survival prognosis ( HR=1.901, P<0.001) compared with the low ratio. We further confirmed that the abundance of collagen in tumor was significantly increased compared with the normal adjacent tissues by the Masson staining [41.0 (14.0-75.0) vs.15.0 (3.0-57.0), P<0.001] and the Sirius red staining [42.5 (10.0-90.0) vs.10.0 (2.5-60.0), P<0.001] on 30 ccRCC tissues and adjacent normal tissues. Based on the Masson staining, we found that high collagen abundance in tumor tissue was associated with more G 3-4 grade of tumor compared with low collagen abundance (38.5% vs.21.3%, OR=2.316, 95% CI 1.146-4.681, P=0.023). Kaplan-Meier survival curve showed that higher collagen abundance was associated with a worse overall survival prognosis in ccRCC ( HR=2.630, P=0.007). However, incomplete fibrous pseudocapsule was associated with a worse overall survival prognosis ( HR=11.140, P<0.001). Conclusions:In ccRCC, intratumoral collagen fiber level was overexpressed. High intratumoral collagen level and incomplete fibrous pseudocapsule may indicate a poor overall survival prognosis.
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At present, the diagnosis and recurrence monitoring of bladder cancer are mainly achieved by invasive cystoscopy, while non-invasive urine diagnostic techniques such as exfoliative cytology, fluorescence in situ hybridization (FISH) and bladder tumor antigen (BTA) are not highly sensitive and/or specific, and are prone to missed diagnosis or misdiagnosis. Therefore, the development of high-sensitivity non-invasive diagnostic techniques for bladder cancer can improve the current status of diagnosis and treatment while reducing the suffering of patients. Recently, the internationally renowned journal "Journal of Clinical Investigation" published the research results of Chinese scholars, Jian Huang/Tianxin Lin and their team who have developed a non-invasive technique for bladder cancer based on urine DNA methylation, which is significantly more sensitive than that clinically common-used technology such as exfoliative cytology and FISH. In particular, it has shown significant advantages in the diagnosis of early, minimal, residual or recurrent bladder cancer, which significantly improves the diagnosis rate of bladder cancer. This technique is expected to reduce the use of cystoscopy and provides new methods for precise diagnosis and treatment of bladder cancer. This article elaborates the current status of non-invasive diagnosis technology for bladder cancer, the advantages of urine DNA methylation detection in the diagnosis of bladder cancer, as well as the trend of development for urine non-invasive diagnosis.
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Objective To establish computer assisted virtual ureteroscopy (VU) through data from computerized tomography urography (CTU) of patients with renal stones and make validation of effectiveness.Methods From June of 2015 to January of 2016,23 cases of renal stones cases was selected by 5 experts in 3 different centers.There were 21 unbilateral cases and 2 bilateral cases.The age ranged from 31 to 79(54.7 ± 12.5).Mean stone burden was (19.0 ± 6.2) mm.Stone number ranged from 1 to 5 (2.7 ± 1.2).VU generation was accomplished by specialized software (Crusher) with incorporating CTU data.After patientspecific VUs were presented to the experts,and the FURS surgeries were all finished successfully,face and content validations about VU using modified Likert questionnaire ordinal 10-point rating scales were made.20 trainee were selected to do the flexible ureteroscopy lithotripsy with assistance of VU.After observation of CTU and VU,the numbers of renal calyces and stones found by the experts and trainees were recorded.The statistical analysis were made before and after observation of VU between the experts and trainees.Result Face and content validation of VU:overall usefulness 7.6 ± 0.5,graphics 7.6 ± 0.5,intrarenal collecting system 8.4 ± 0.5,stone details 8.4 ± 0.5,usefulness in surgical planning and training 8.0 ± 0.7.Significant improvement was found when the trainees doing the surgery with the help of VU.Compared with using CTU only,VU could help the trainees had better understanding of intrarenal structure and stone information [the number of calyces (16.7 ±3.7)vs.(24.6 ± 1.8),P ＜0.001;the number of stones (4.9 ± 1.4)vs.(8.2 ± 1.3),P ＜0.001].Before observation of VU,trainees found much fewer calyces and stones compared with experts (P =0.004 and P ＜ 0.001 respectively).However,this gap disappeared after VU observation (P =0.327 and 0.292 respectively).Conclusions Establishing computer assisted VU through CTU data from renal stone patients is feasible and rapid.VU can significantly improve trainee's view of intrarenal collecting system and stone information before practicing FURS.
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Objective:To investigate the expression of MEK/ERK signaling pathways in renal cell car-cinoma with bone metastasis,and to analyze the differences of expressions of VEGFR-2,MEK,ERK on the primary and metastasis tissue and its mechanism.Methods:The tissue samples were obtained from 7 renal cell carcinoma patients kindly provided by Department of Urology,Peking University People’s Hos-pital from January 1,2009 to January 1,2010.The expression of MEK/ERK signaling pathways was de-tected in the 7 renal cell carcinoma patients`primary and matched metastatic tissues with ICH,The anti-body concentrations were 1 ∶200,1 ∶25,and 1 ∶250,respectively.The mutation of the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the se-cond exon of the MEK1 gene were detected with PCR.Results:The expression intensities of VEGFR-2, MEK,and ERK were measured by H-score [intensity (1,2,3,or 4)multiplied by the distribution (%)].VEGFR-2,MEK,and ERK expressions were divided into 3 groups according to the positive dis-tribution of the tumor cells:1,0 -5%;2,6% -50%;and 3,>50%,To assess intratumor heteroge-neity,three distinct microscopic fields (×200)from each specimen were used to evaluate the expres-sions,Subsequently,the scores were averaged to obtain a single concatenated score for each tissue. VEGFR-2,MEK,and ERK expressions were assessed by 2 independent pathologists who were blinded to the clinicopathological data.The data were expressed as the mean value of the triplicate experiments.The expressions of MEK,and ERK were higher in the metastatic tissues than in the matched RCC tissues (6.10 ±4.10 vs.1.33 ±0.51,P =0.015;9.10 ±2.24 vs.4.43 ±2.84,P =0.021 )while the ex-pression of VEGFR-2 was not different between the primary and metastatic tissues (P =0.901).No mu-tation was detected on the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the second exon of the MEK1 gene.Conclusion:MEK/ERK signa-ling pathways may play an important role in the metastasis and the resistance of sunitinib in RCC patients with bone metastasis.
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Objective:To investigate the change of biological characteristics after stable knockdown of CKLF-like MARVEL transmembrane domain containing 3 (CMTM3)expression in PC3 by lentivirus shRNA and to reveal new therapeutic targets.Methods:The research includes two groups:sh393 is the experimental group in which CMTM3 is knocked down in PC3 cell line;shN is the control group in which CMTM3 is negatively knocked down.The expression of CMTM3 was detected by Western blot.The mi-gration ability of PC3 after stable knockdown was detected by Transwell and Wound healing assay.The invasion ability of PC3 was detected by Matrigel assay.Results were obtained from at least three indivi-dual experiments.Results:The expression of CMTM3 in sh393 group is significant lower than shN group (0.004 0 ±0.000 4 vs.0.490 0 ±0.055 7,P <0.001)detected by Western blot.It also had statistical significance in Matrigel assays (248.6 ±4.5 vs.113.0 ±3.3),Transwell (203.6 ±1.9 vs.103.0 ± 1.2)and Wound healing assays (95.0 ±2.9 vs.33.0 ±1.5)that knockdown of CMTM3 promoted mi-gration,and invasion of PC3 cells in vitro (P <0.001).Conclusion:Negative correlation exists between the stable knockdown of CMTM3 and change of biological characteristics in PC3 cells,and knocking down CMTM3 affects migration,and invasion ability in PC3 cells.
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Objective To investigate the expression of CKLF-like MARVEL transmembrane (CMTM)domain-containing family in clear cell renal cell carcinoma(ccRCC)and its significance.Methods Seventy-five samples of ccRCC were collected,including 50 males and 25 females,mean age (59 ± 10)years.There were 34 cases in clinical stage Ⅰ,23 cases in stage Ⅱ,14 cases in stage Ⅲ and 4 cases in stage Ⅳ.The pathological differentiation was 3 cases of grade Ⅰ,1 case of grade Ⅰ-Ⅱ,35 cases of grade Ⅱ,10 cases of grade Ⅱ-Ⅲ,18 cases of grade Ⅲ,3 cases of grade Ⅲ-Ⅳ and 5 cases of grade Ⅳ.The expression of CMTM3 and CMTM5 proteins in 75 cases of ccRCC and corresponding adjacent normal kidney tissues was detected by tissue microarray and immunohistochemistry.Results The positive expression rate of CMTM3 and CMTM5 was 98.7％,97.3％ in the adjacent normal kidney tissues,and 44.0％,68.0％ in ccRCC tissues(P ＜ 0.05).The expression of CMTM3 and CMTM5 had no correlation with the gender,age,clinical staging and pathological differentiation(P ＞ 0.05).Conclusion CMTM3 and CMTM5 could be ccRCC suppressor genes,and their mutation or methylation might be an early event in the carcinogenesis of ccRCC,thus promise them to be potential biomarkers in early diagnosis.