Multi-omics and immunogenomics analysis revealed PFKFB3 as a targetable hallmark and mediates sunitinib resistance in papillary renal cell carcinoma: in silico study with laboratory verification.

Glycolysis-related metabolic reprogramming is a central hallmark of human cancers, especially in renal cell carcinoma. However, the regulatory function of glycolytic signature in papillary RCC has not been well elucidated. In the present study, the glycolysis-immune predictive signature was constructed and validated using WGCNA, glycolysis-immune clustering analysis. PPI network of DEGs was constructed and visualized. Functional enrichments and patients' overall survival were analyzed. QRT-PCR experiments were performed to detect hub genes' expression and distribution, siRNA technology was used to silence targeted genes; cell proliferation and migration assays were applied to evaluate the biological function. Glucose concentration, lactate secretion, and ATP production were measured. Glycolysis-Immune Related Prognostic Index (GIRPI) was constructed and combined analyzed with single-cell RNA-seq. High-GIRPI signature predicted significantly poorer outcomes and relevant clinical features of pRCC patients. Moreover, GIRPI also participated in several pathways, which affected tumor immune microenvironment and provided potential therapeutic strategy. As a key glycolysis regulator, PFKFB3 could promote renal cancer cell proliferation and migration in vitro. Blocking of PFKFB3 by selective inhibitor PFK-015 or glycolytic inhibitor 2-DG significantly restrained renal cancer cells' neoplastic potential. PFK-015 and sunitinib could synergistically inhibit pRCC cells proliferation. Glycolysis-Immune Risk Signature is closely associated with pRCC prognosis, progression, immune infiltration, and therapeutic response. PFKFB3 may serve as a pivotal glycolysis regulator and mediates Sunitinib resistance in pRCC patients.

Quantitative Cerebral Blood Volume Image Synthesis from Standard MRI Using Image-to-Image Translation for Brain Tumors.

Background Cerebral blood volume (CBV) maps derived from dynamic susceptibility contrast-enhanced (DSC) MRI are useful but not commonly available in clinical scenarios. Purpose To test image-to-image translation techniques for generating CBV maps from standard MRI sequences of brain tumors using the bookend technique DSC MRI as ground-truth references. Materials and Methods A total of 756 MRI examinations, including quantitative CBV maps produced from bookend DSC MRI, were included in this retrospective study. Two algorithms, the feature-consistency generative adversarial network (GAN) and three-dimensional encoder-decoder network with only mean absolute error loss, were trained to synthesize CBV maps. The performance of the two algorithms was evaluated quantitatively using the structural similarity index (SSIM) and qualitatively by two neuroradiologists using a four-point Likert scale. The clinical value of combining synthetic CBV maps and standard MRI scans of brain tumors was assessed in several clinical scenarios (tumor grading, prognosis prediction, differential diagnosis) using multicenter data sets (four external and one internal). Differences in diagnostic and predictive accuracy were tested using the z test. Results The three-dimensional encoder-decoder network with T1-weighted images, contrast-enhanced T1-weighted images, and apparent diffusion coefficient maps as the input achieved the highest synthetic performance (SSIM, 86.29% ± 4.30). The mean qualitative score of the synthesized CBV maps by neuroradiologists was 2.63. Combining synthetic CBV with standard MRI improved the accuracy of grading gliomas (standard MRI scans area under the receiver operating characteristic curve [AUC], 0.707; standard MRI scans with CBV maps AUC, 0.857; z = 15.17; P < .001), prediction of prognosis in gliomas (standard MRI scans AUC, 0.654; standard MRI scans with CBV maps AUC, 0.793; z = 9.62; P < .001), and differential diagnosis between tumor recurrence and treatment response in gliomas (standard MRI scans AUC, 0.778; standard MRI scans with CBV maps AUC, 0.853; z = 4.86; P < .001) and brain metastases (standard MRI scans AUC, 0.749; standard MRI scans with CBV maps AUC, 0.857; z = 6.13; P < .001). Conclusion GAN image-to-image translation techniques produced accurate synthetic CBV maps from standard MRI scans, which could be used for improving the clinical evaluation of brain tumors. Published under a CC BY 4.0 license. Supplemental material is available for this article. See also the editorial by Branstetter in this issue.

Semi-Supervised Standard-Dose PET Image Generation via Region-Adaptive Normalization and Structural Consistency Constraint.

Positron Emission Tomography (PET) is an important nuclear medical imaging technique, and has been widely used in clinical applications, e.g., tumor detection and brain disease diagnosis. As PET imaging could put patients at risk of radiation, the acquisition of high-quality PET images with standard-dose tracers should be cautious. However, if dose is reduced in PET acquisition, the imaging quality could become worse and thus may not meet clinical requirement. To safely reduce the tracer dose and also maintain high quality of PET imaging, we propose a novel and effective approach to estimate high-quality Standard-dose PET (SPET) images from Low-dose PET (LPET) images. Specifically, to fully utilize both the rare paired and the abundant unpaired LPET and SPET images, we propose a semi-supervised framework for network training. Meanwhile, based on this framework, we further design a Region-adaptive Normalization (RN) and a structural consistency constraint to track the task-specific challenges. RN performs region-specific normalization in different regions of each PET image to suppress negative impact of large intensity variation across different regions, while the structural consistency constraint maintains structural details during the generation of SPET images from LPET images. Experiments on real human chest-abdomen PET images demonstrate that our proposed approach achieves state-of-the-art performance quantitatively and qualitatively.

Effect of Radical Laparoscopic Surgery and Conventional Open Surgery on Surgical Outcomes, Complications, and Prognosis in Elderly Patients with Bladder Cancer.

Background: Bladder cancer is a common malignant tumor of the urinary system in the clinic. It has multiple lesions, easy recurrence, easy metastasis, poor prognosis, and high mortality. Objective: The aim of this study is to investigate the impact of laparoscopic radical cystectomy (LRC) and open radical cystectomy (ORC) on the surgical outcome, complications, and prognosis of elderly patients with bladder cancer. Materials and Methods: One hundred elderly bladder cancer patients who underwent surgery in our hospital from June 2019 to June 2021 were selected for the retrospective study and were divided into 50 cases each in the ORC group and the LRC group according to the different surgical methods. The ORC group was treated with ORC, and the LRC group implemented LRC treatment. The differences in surgery, immune function, recent clinical outcomes, and complications between the two groups were observed and compared. Results: The mean operative time, mean intraoperative bleeding, intraoperative and postoperative transfusion rate, and transfusion volume of patients in the LRC group were statistically significant when compared to the ORC group. The differences in the meantime to resume eating, time to get out of bed, mean number of days in hospital after surgery, and the amount of postoperative numbing analgesics used by patients in the LRC group after surgery were statistically significant compared to the ORC group (P < 0.05). There was no statistically significant difference in the comparison of immune function between the two groups before surgery (P > 0.05), while the comparison of CD8+ and B cells 1 week after surgery of the LRC group was significantly better than that of the ORC group (P < 0.05), and the operation time of the LRC group was longer than that of the ORC group (P < 0.05). Statistical analysis of postoperative complications showed that the overall incidence of postoperative complications in the LRC group was significantly lower than that in the ORC group (16.67% vs. 46.67%) (P < 0.05). Conclusion: LRC has less surgical trauma and intraoperative bleeding, faster postoperative recovery, and fewer postoperative complications, providing some reference for clinical surgery for elderly bladder cancer patients.

Demethylase FTO inhibits the development of prostate cancer by upregulating EGR2 expression in an m6A manner.

Fat mass and obesity-associated protein (FTO) is a demethylase and plays a vital role in various cancers. However, the regulation mechanism of FTO in prostate cancer (PCa) remains unclear. This study aimed to elucidate the mechanism of FTO in PCa. The function and mechanism of FTO-mediated in PCa were determined by gain-of-function assays and RNA-seq. We found that FTO expression in PCa tissues and two PCa cell lines were significantly lower than that in adjacent tissues and normal cell line. PCa cells after overexpression of FTO showed a significant lower in proliferation, migration, and invasion capabilities. RNA-seq displayed that FTO overexpression altered transcriptome landscape in Du145 and PC-3 cells, particularly upregulating EGR2 expression. FTO overexpression induced differential expression genes, including MYLK2, DNA2, CDK, and CDC (6, 7, 20, 25, and 45), which were mainly enriched in adjustment of cell cycle and growth pathways. Furthermore, FTO overexpression significantly reduced the EGR2 methylation level. Arresting the proliferation, migration, and invasion of Du145 cells induced by FTO overexpression was significantly rescued by EGR2 knockdown. FTO overexpression also significantly inhibited tumor growth and promoted EGR2 protein expression. Taken together, FTO suppresses PCa progression by regulating EGR2 methylation. We uncovered a novel regulatory mechanism of FTO in PCa and provide a new potential therapeutic target for PCa.

bpMRI and mpMRI for detecting prostate cancer: A retrospective cohort study.

Background: We aimed to compare the detection rates of prostate cancer (PCa) and clinically significant prostate cancer(csPCa) by biparametric (bp-) and multiparameter magnetic resonance imaging (mpMRI). Materials and Methods: A total of 699 patients who underwent transperineal prostate biopsy in the Department of Urology, the Second Affiliated Hospital of Nantong University from January 2018 to December 2021 were retrospectively reviewed. Multivariate analysis was used to explore the influencing factors associated with the detection rates of PCa and csPCa. According to MRI examination before biopsy, the patients were divided into bpMRI group and mpMRI group. The detection rates of PCa and csPCa by bpMRI and mpMRI were compared. Furthermore, stratified analysis was performed for patients in these two groups to compare the detection rates of PCa and csPCa at different tPSA intervals, different prostate volume (PV) intervals and different PI-RADS V2 scores. Results: A total of 571 patients were finally analyzed in this study after exclusion, and the overall detection rate of PCa was 54.5%. Multivariate analysis showed that patient age, tPSA level, prostate volume and PI-RADS V2 score were independent risk factors affecting the detection rates of PCa and csPCa. The detection rates of PCa and csPCa by bpMRI and mpMRI were comparable (51.3% vs. 57.9%, 44.0% vs. 48.0%, both P > 0.05), with no statistical significance. In the tPSA 10-20 ng/ml interval, the detection rates of PCa (59.72% vs. 40.35%, P = 0.011) and csPCa (51.39% vs. 28.82%, P = 0.005) by mpMRI were significantly higher than those by bpMRI, while in other tPSA interval (tPSA < 4 ng/ml, 4-10 ng/ml, 20-100 ng/ml), different PVs (≤30 ml, 30-60 ml, >60 ml) and different PI-RADS V2 scores (3, 4, and 5), the detection rates of PCa and csPCa were comparable between the two groups. Conclusion: For detecting PCa and csPCa, bpMRI and mpMRI had similar diagnostic efficacies, whereas mpMRI detected more PCa and csPCa in the tPSA interval of 10-20 ng/ml.

Diagnosis of Liver Neoplasms by Computational and Statistical Image Analysis.

BACKGROUND: Distinguishing well-differentiated hepatocellular carcinoma (WD-HCC), hepatocellular adenoma (HA) and non-neoplastic liver tissue (NNLT) solely on morphology is often challenging. The purpose of this study was to evaluate the use of computational image analysis to distinguish WD-HCC, HA and NNLT. METHODS: Seventy-seven cases comprising of WD-HCC (n = 26), HA (n = 23) and NNLT (n = 28) were retrieved and reviewed. A total of 485 hematoxylin and eosin (H&E) photomicrographs (× 400, 0.09 µm2) of WD-HCC (n = 183), HA (n = 173), NNLT (n = 129) and nine whole-slide scans (three of each diagnosis) were obtained, color deconvoluted and digitally transformed. Quantitative data including nuclear density, nuclear sphericity, nuclear perimeter, and nuclear eccentricity from each image were acquired. The data were analyzed by one-way analysis of variance (ANOVA) with Tukey post hoc test, followed by unsupervised and supervised (Chi-square automatic interaction detection (CHAID)) cluster analysis. RESULTS: Unsupervised cluster analysis identified three well defined clusters of WD-HCC, HA and NNLT. Employing the four most discriminating nuclear features, supervised analysis was performed on a training set of 383 images, and validated on the remaining 102 test images. The analysis identified WD-HCC (sensitivity 100%, specificity 98%), HA (sensitivity 71%, specificity 85%) and NNLT (sensitivity 70%, specificity 86%). An analysis of whole-slide images identified WD-HCC with sensitivity and specificity of 100%. CONCLUSIONS: We have successfully demonstrated that computational image analysis of nuclear features can differentiate WD-HCC from non-malignant liver with high accuracy, and can be used to assist in the histopathological diagnosis of hepatocellular carcinoma.

p38gamma overexpression promotes renal cell carcinoma cell growth, proliferation and migration.

Recent studies have proposed that p38gamma (p38γ) might be critically involved in tumorigenesis and cancer progression. Its expression and potential functions in human renal cell carcinoma (RCC) are studied here. We show that p38γ mRNA and protein levels are upregulated in human RCC tissues, as compared to its levels in the surrounding normal renal tissues. p38γ upregulation was also detected in established (786-O line) and primary human RCC cells. Functional studies in 786-O cells and primary human RCC cells demonstrated that p38γ silencing (by targeted shRNAs) or CRISPR/Cas9-mediated p38γ knockout (KO) potently inhibited cell growth, viability, proliferation and migration. Furthermore, p38γ shRNA or KO in RCC cells decreased retinoblastoma (Rb) phosphorylation and downregulated cyclin E1/A expression. Additionally, significant apoptosis activation was detected in p38γ-silenced and p38γ-KO RCC cells. Contrarily, ectopic overexpression of p38γ facilitated cell growth, viability, proliferation and migration in RCC cells. Taken together, we show that p38γ overexpression promotes RCC cell growth, proliferation and migration. p38γ could be a novel therapeutic target for human RCC.

BMI and serum lipid parameters predict increasing risk and aggressive prostate cancer in Chinese people.

OBJECTIVES: To determine if obesity and serum lipid parameters are associated with increased risk and more aggressive prostate cancer in Chinese population. MATERIALS AND METHODS: We conducted a retrospective cohort analysis including 3102 patients. Kruskal-Wallis test for continuous variables and the chi-squared tests for categorical variables were used for univariate comparison of the differences in patient characteristics across BMI categories between different groups. Odds ratios (OR) and 95% confidence intervals (CI) were estimated for the association between prostate cancer and the various patient characteristics. Multivariable Cox proportional hazards regression was performed to assess the risk of prostate cancer recurrence. RESULTS: 974 consecutive men were diagnosed as prostate cancer and 700 patients subsequently received radical prostatectomy immediately, and 1031 patients were pathologically diagnosed as biopsy negative. The level of low-density-lipoprotein cholesterol (LDL-c) and total cholesterol was significantly higher and the high-density-lipoprotein cholesterol (HDL-c) level is much lower in prostate cancer patients. Patients with low level of HDL-c, who subsequently received radical prostatectomy, had increased risk of high risk disease. In addition, patients with normal weight were less likely to develop a biochemical recurrence. Combined analysis revealed that obese patients had significantly higher rates of PSA recurrence over time than nonobese patients. CONCLUSIONS: In our study, lipid parameters are supposed to be associated with prostate cancer risk and aggressiveness. Obese men are at increased risk of PSA recurrence after radical prostatectomy.

Characteristics of prostate cancer detection rate (PCDR) in Chinese Han population under different prostate biopsy methods.

We analyzed the improvement of prostate cancer detection rate (PCDR) in Chinese Han population and summarized the characteristics of prostate cancer (PCa) with the advancement of prostate biopsy technologies. From March 1999 to March 2015, 3762 patients underwent the systematic 6-, 8- or 13-core biopsy, guided by finger or transrectal ultrasound (TRUS) at our center. The PCDR under different PSA intervals and different biopsy methods were analyzed. The trends of PSA level, age and Gleason score of PCa patients were summarized. The PCDR of finger-guided 6- and 8-core biopsies were 30.8% (340/1103) and 36.7% (147/401) respectively. In 2258 patients with TRUS-guided 13-core biopsies, 992 (43.9%) were diagnosed as PCa, higher than that with finger-guided biopsies (43.9% vs. 32.4%, p < 0.001). The PCDR of prostate peripheral zone was higher than that of medial zone (37.5% vs. 31.4%, p < 0.001). Interestingly, the PCDR of extra 13th core was higher than the mean positive rate of other 12 cores (70.7% vs. 56.0%, p < 0.001). The systematic 13-core prostate biopsy guided by TRUS is safe, effective, and economic for PCa diagnosis in developing countries like China. The extra 13th core biopsy is beneficial to increase the PCDR.

The Association between Abnormal Long Noncoding RNA MALAT-1 Expression and Cancer Lymph Node Metastasis: A Meta-Analysis.

Previous studies have investigated that the expression levels of MALAT-1 were higher in cancerous tissues than matched histologically normal tissues. And, to some extent, overexpression of MALAT-1 was inclined to lymph node metastasis. This meta-analysis collected all relevant articles and explored the association between MALAT-1 expression levels and lymph node metastasis. We searched PubMed, EmBase, Web of Science, Cochrane Library, and OVID to address the level of MALAT-1 expression in cancer cases and noncancerous controls (accessed February 2015). And 8 studies comprising 696 multiple cancer patients were included to assess this association. The odds ratio (OR) and its corresponding 95% confidence interval (CI) were calculated to assess the strength of the association using Stata 12.0 version software. The results revealed there was a significant difference in the incidence of lymph node metastasis between high MALAT-1 expression group and low MALAT-1 expression group (OR = 1.94, 95% CI 1.15-3.28, P = 0.013 random-effects model). Subgroup analysis indicated that MALAT-1 high expression had an unfavorable impact on lymph node metastasis in Chinese patients (OR = 1.87, 95% CI 1.01-2.46). This study demonstrated that the incidence of lymph node metastasis in patients detected with high MALAT-1 expression was higher than that in patients with low MALAT-1 expression in China.

Developing a Correct System to Evaluate the Accuracy of Gleason Score in Prostate Cancer of Chinese Population.

INTRODUCTION: A study was conducted to develop a new correct system to improve the overall rate of Gleason sum concordance between biopsy and final pathology. MATERIALS AND METHODS: A total of 592 consecutive patients who had undergone transrectal ultrasound-guided prostate biopsy and radical prostatectomy were evaluated during the first stage. Age, PSA, PSA density (PSAD), biopsy cores, positive cores, prostate volume, positive core rate (PCR), core volume rate (CVR) and digital rectal examination findings were considered predictive factors. A multiple logistic regression analysis involving a backward elimination selection procedure and linear regression analysis involving a stepwise procedure were applied to select independent predictors. RESULTS: Positive cores, PCR, CVR and PSAD were included in our assessing credibility model in the first stage. A significantly higher area under the receiver-operating curve was obtained in our model compared with CVR alone (0.641 vs. 0.517). In the second stage, patients with credibility of pre-operative Gleason score <0.388 were subjected to further evaluation. Compared with the 2 statuses, the rate of overall concordance was significantly increased (60.3 vs. 50.2%, p = 0.002). CONCLUSIONS: We developed a follow-up strategy based on the new and correct system, which represents an important consideration procedure when clinicians make decisions with regard to treatment plans.

Developing a model for forecasting Gleason score ≥7 in potential prostate cancer patients to reduce unnecessary prostate biopsies.

PURPOSE: The diagnosis of Gleason score (GS) ≥7 with distinction from GS < 7 remains a difficult problem instructing clinical decisions. Moreover, the present wide application of prostate biopsy to increase prostate cancer detection rate might cause unnecessary and excessive examination or treatment. Therefore, a risk assessment model for forecasting GS ≥ 7 in potential prostate cancer patients was established to reduce unnecessary prostate biopsies. METHODS: Patients (n = 981; September 2009 to January 2013) who underwent trans-rectal ultrasound (TRUS)-guided core prostate biopsy were retrospectively evaluated in the first stage of the study. Age, prostate-specific antigen (PSA), free PSA (fPSA), the free/total PSA ratio (f/t), prostate volume (PV), PSA density (PSAD), digital rectal examination (DRE) findings (texture, nodules) and B-ultrasound detection results (normal or abnormal, presence of hypoechoic mass or microcalcification) were considered as potential predictive factors. After multiple logistic regression analysis, independent variables used to build a nomogram were selected using a backward elimination selection procedure. Then, a model to forecast GS ≥ 7 was designed for potential prostate cancer patients. In the second stage of the study, 410 cases (January 2013 to March 2015) were subsequently evaluated using our model for prostate biopsies, and the outcomes of biopsies were compared between the two stages. RESULTS: PSA, DRE texture, DRE nodules and B-ultrasound results were finally brought into our nomogram; a obviously greater area under the receiver operating characteristic (ROC) curve was obtained for the model than utilizing PSA, fPSA or PSAD alone (0.831 vs. 0.803, 0.770, 0.780 separately). We thereafter sought the best cutoff value in the ROC curve at 0.87, which provided sensitivity as high as 90%. Meanwhile, the specificity was 45.8%, which was much higher than the specificity of PSA, fPSA and PSAD at the same sensitivity level (37.7, 24.6 and 35.2%, respectively). In the first stage, the detection rate of GS ≥ 7 in the high-risk group was significantly higher than in the low-risk group (80.3 vs. 35.0%, p < 0.001). Furthermore, in the second stage, with the application of the new model associated with our former models, the rate of GS ≥ 7 was improved from 71.0 (697/981) to 79.2% (267/337) (p = 0.003). CONCLUSIONS: The model for forecasting GS ≥ 7 is effective, which could reduce unnecessary prostate biopsies without delaying patients' diagnoses and treatments.

Overexpression of long non-coding RNA LOC400891 promotes tumor progression and poor prognosis in prostate cancer.

Tumor recurrence and metastasis remain the major obstacles for the successful treatment of patients diagnosed with prostate cancer (PCa). In recent years, long non-coding RNAs (lncRNAs) have been considered as key regulators of tumor behavior. In this study, we investigated the biological role and clinical relevance of the lncRNA LOC400891 in prostate cancer. Using of lncRNAs expression chips screening and the biological analysis, we found the target lncRNA (LOC400891). Moreover, the expression levels of lncRNA LOC400891 in PCa tissues and cell lines were evaluated by quantitative real-time PCR (qRT-PCR), and its association with biochemical recurrence-free survival of patients was analyzed by statistical analysis. Furthermore, the effect of LOC400891 on proliferation, migration, and invasion was studied in PCa cells. We found that the expression level of LOC400891 was higher in PCa tissues and cells compared to adjacent non-tumor tissues and normal prostate stromal immortalized cells WPMY-1. The patients with higher LOC400891 expression had an advanced clinical features and a shorter biochemical recurrence-free survival time than those with lower LOC400891 expression. Furthermore, multivariate analysis showed that the status of LOC400891 expression was an independent predictor of biochemical recurrence-free survival in PCa. We also found that knockdown of LOC400891 could inhibit cell proliferation, migration, and invasion in vitro study. Our data suggested that lncRNA LOC400891 was a novel molecule involved in PCa progression, which provided a potential prognostic biomarker and therapeutic target.

Genetic Variants in Caveolin-1 and RhoA/ROCK1 Are Associated with Clear Cell Renal Cell Carcinoma Risk in a Chinese Population.

BACKGROUND: The RhoA/ROCK pathway and Caveolin-1 (Cav-1) participate in the process of tumorigenesis in numerous types of cancer. Up-regulation of RhoA/ROCK and Cav-1 expression is considered to be associated with the development and progression of clear cell renal cell carcinoma (ccRCC). We investigated the association between genetic variations of RhoA/ROCK and Cav-1 and the risk of ccRCC in the Chinese population. METHODS: Between May 2004 and March 2014, a total of 1,248 clear cell renal cell carcinoma cases and 1,440 cancer-free controls were enrolled in this hospital-based case-control study. Nine SNPs in RhoA/ROCK and Cav-1 were genotyped using the TaqMan assay. RESULT: We found two SNPs (Cav-1 rs1049334 and ROCK1 rs35996865) were significantly associated with the increasing risk of ccRCC (P = 0.002 and P < 0.001 respectively). The analysis of combined risk alleles revealed that patients with 2-4 risk alleles showed a more remarkable growth of ccRCC risk than the patients with 0-1 risk alleles(OR = 1.66, 95%CI = 1.31-2.11, P < 0.001). Younger subjects (P = 0.001, OR = 1.83, 95%CI = 1.30-2.57), higher weight subjects (P = 0.001, OR = 1.76, 95%CI = 1.25-2.47), female subjects (P = 0.007, OR = 1.75, 95% CI = 1.17-2.62), nonsmokers (P < 0.001, OR = 1.67, 95%CI = 1.26-2.23), drinkers (P = 0.025, OR = 1.75, 95% CI = 1.07-2.85), subjects with hypertension (P = 0.025, OR = 1.75, 95% CI = 1.07-2.85) and diabetes (P = 0.026, OR = 4.31, 95% CI = 1.19-15.62) showed a stronger association between the combined risk alleles and the risk of ccRCC by using the stratification analysis. Furthermore, we observed higher Cav-1 mRNA levels in the presence of the rs1049334 A allele in normal renal tissues. CONCLUSION: Our results indicate that the two SNPs (Cav-1 rs1049334 and ROCK1 rs35996865) and genotypes with a combination of 2-4 risk alleles were associated with the risk of ccRCC. The functional SNP rs1049334 may affect the risk of ccRCC by altering the expression of Cav-1 and the relevance between the risk effects and the functional impact of this polymorphism needs further validation.