Integrative analysis of deoxyribonuclease 1-like 3 as a potential biomarker in renal cell carcinoma.

Background: Clear cell renal cell carcinoma (ccRCC), the most common subtype of renal cell carcinoma (RCC), is insensitive to radiotherapy and chemotherapy after surgery. Deoxyribonuclease 1-like 3 (DNASE1L3), an endonuclease that cleaves both membrane-encapsulated single- and double-stranded DNA, suppresses cell cycle progression, proliferation and metabolism in hepatocellular carcinoma cells. There is currently no established link between DNASE1L3 and RCC inhibition. We are gonging to explored the mechanism underlying the relationship between DNASEL1L3 and RCC. Methods: RNA sequencing data for RCC tissue and peritumoral tissue were downloaded from The Cancer Genome Atlas database and analyzed. The expression levels of DNASE1L3 in RCC and normal samples were verified using the Gene Expression Omnibus (GEO) database, Human Protein Atlas database and western blotting. The role and potential mechanism of DNASE1L3 were investigated by analysis of immune-related databases and wound healing, invasion, cell counting kit 8 and immunofluorescence assays. Results: We revealed that DNASE1L3 expression was downregulated in RCC group compared with control group [The Cancer Genome Atlas (TCGA): 7.98 vs. 10.87, P<0.001]. Meanwhile, DNASE1L3 expression correlated with the clinical characteristics of patients. Patients with low DNASE1L3 expression had worse survival (P<0.001) and larger (r=-0.32, P<0.001) and heavier tumors (r=-0.17, P<0.001). DNASE1L3 overexpression inhibited the proliferation (786-O: 0.135±0.014 vs. 0.322±0.027, P<0.001) and invasion (786-O: 1,479±134 vs. 832±67, P<0.05) of RCC cells. The expression of DNASE1L3 was significantly correlated with the tumor immune microenvironment and drug sensitivity in ccRCC. Moreover, the level of the key phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway protein P-AKT was decreased in the group of cells transfected with DNASE1L3. Conclusions: This study strongly suggest that DNASE1L3 may be a promising potential biomarker for the diagnosis and treatment of ccRCC patients.

GPD1L inhibits renal cell carcinoma progression by regulating PINK1/Parkin-mediated mitophagy.

Few approaches have been conducted in the treatment of renal cell carcinoma (RCC) after nephrectomy, resulting in a high mortality rate in urological tumours. Mitophagy is a mechanism of mitochondrial quality control that enables selective degradation of damaged and unnecessary mitochondria. Previous studies have found that glycerol-3-phosphate dehydrogenase 1-like (GPD1L) is associated with the progression of tumours such as lung cancer, colorectal cancer and oropharyngeal cancer, but the potential mechanism in RCC is still unclear. In this study, microarrays from tumour databases were analysed. The expression of GPD1L was confirmed by RT-qPCR and western blotting. The effect and mechanism of GPD1L were explored using cell counting kit 8, wound healing, invasion, flow cytometry and mitophagy-related experiments. The role of GPD1L was further confirmed in vivo. The results showed that GPD1L expression was downregulated and positively correlated with prognosis in RCC. Functional experiments revealed that GPD1L prevented proliferation, migration and invasion while promoting apoptosis and mitochondrial injury in vitro. The mechanistic results indicated that GPD1L interacted with PINK1, promoting PINK1/Parkin-mediated mitophagy. However, inhibition of PINK1 reversed GPD1L-mediated mitochondrial injury and mitophagy. Moreover, GPD1L prevented tumour growth and promoted mitophagy by activating the PINK1/Parkin pathway in vivo. Our study shows that GPD1L has a positive correlation with the prognosis of RCC. The potential mechanism involves interacting with PINK1 and regulating the PINK1/Parkin pathway. In conclusion, these results reveal that GPD1L can act as a biomarker and target for RCC diagnosis and therapy.

The Effects of circ_000558/miR-1225-5p/ARL4C on Regulating the Proliferation of Renal Cell Carcinoma Cells.

Renal cell carcinoma (RCC) is one of the top ten tumors over the world. RCC is not sensitive to radiotherapy and chemotherapy. Therefore, it is necessary to find new targets for the treatment. CircRNAs are a special type of noncoding RNAs, which play important roles in many types of cancer. In this study, we found circ_000558 was upregulated in RCC cells, and it elevated the proliferation ability of RCC cells. The relationship between miR-1225-5p and circ_000558 or ARL4C was predicted via circBank and circular RNA interactome and confirmed by dual-luciferase reporter assay. Then, the effects of circ_000558/miR-1225-5p/ARL4C on RCC cell proliferation and apoptosis were assessed by CCK-8 assay. The results revealed that the knockdown of ARL4C significantly reduced RCC cell proliferation and overexpression of circ_000558 could significantly induce RCC cell proliferation after miR-1225-5p treatment further promoted the inhibitory ability of ARL4C knockdown. Overall, our study suggested that circ_000558/miR-1225-5p/ARL4C network was related to the RCC cell proliferation. This finding could provide new targets for the treatment and prognosis of RCC.

Transient Receptor Potential Channel 1 Potentially Serves as a Biomarker Indicating T/TNM Stages and Predicting Long-Term Prognosis in Patients With Renal Cell Carcinoma.

Background: Transient receptor potential channel 1 (TRPC1) regulates the progression of several cancers, but its clinical implication in renal cell carcinoma (RCC) has not been explored yet. This study aimed to investigate the correlation of TRPC1 with clinical characteristics and prognosis in patients with RCC. Methods: Totally, 177 patients with primary RCC who received surgical resection were retrospectively screened. Their tumor and paired adjacent tissue specimens were retrieved to assess TRPC1 mRNA expression using RT-qPCR and TRPC1 protein expression using immunohistochemistry (IHC). Results: Both TRPC1 IHC score and TRPC1 mRNA expression were elevated in RCC tissue than in adjacent tissue (both P < 0.001). Meanwhile, both TRPC1 IHC score and TRPC1 mRNA expression in tumor were associated with higher T stage (both P = 0.02) and TNM stage (P = 0.009, P = 0.003, respectively). However, no correlation was found in tumor TRPC1 IHC score or TRPC1 mRNA expression with other tumor properties (all P > 0.05). Besides, the 3-, 5-, and 7-year overall survival (OS) were 81.4, 68.6, and 60.2%, respectively in patients with high tumor TRPC1 protein, while they were 89.3, 82.7, and 76.7%, respectively in patients with low tumor TRPC1 protein. High (vs. low) TRPC1 protein in the tumor was associated with shorter OS (P = 0.017), while high (vs. low) TRPC1 mRNA in the tumor was not correlated with OS (P = 0.144). By the forward stepwise method, TRPC1 protein expression independently predicted poor OS (P = 0.01, hazard ratio = 2.052). Conclusion: TRPC1 serves as a potential biomarker reflecting tumor features and long-term survival profile in patients with RCC.

CTSV (cathepsin V) promotes bladder cancer progression by increasing NF-κB activity.

Chronic inflammation is positively associated with the development of urinary bladder cancer. However, its detailed regulatory mechanism remains elusive. The quantitative real-time polymerase chain reaction was used to measure mRNA levels of relative genes. The protein levels were monitored by western blotting. Cell proliferation and viability were evaluated by the cell counting Kit 8 (CCK8) and colony formation assays, respectively. The dual-luciferase reporter assay was performed to assay the transcriptional activity. In vivo experiments were implemented in nude mice as well. The TCGA database analysis suggested that the aberrant expression of cathepsin V (CTSV) was related to a poor outcome in bladder cancer patients. CTSV boosted the inflammation reaction, which facilitated the development of bladder cancer. The overexpression of CTSV increased the proliferation and viability of bladder cancer cells. On the contrary, the deletion of CTSV significantly inhibited the proliferation and viability of bladder cancer cells. The tumor repression resulting from CTSV deficiency in vitro was also verified in vivo. Moreover, multiple cancer-associated luciferase screening showed that the overexpression of CTSV triggered the inflammatory signaling pathway, which could be restored by introducing the NF-κB inhibitor. CTSV is upregulated and promotes proliferation through the NF-κB pathway in bladder cancer and may be a potential target in inflammation-associated bladder cancer.

The second short-term warm ischemia after vascular anastomosis did not affect early renal function recovery in renal transplantation: a case report.

Ischemic postconditioning was defined as rapid intermittent interruptions of blood flow in the early phase of reperfusion, which has been found to be protective against renal ischemia-reperfusion injury (IRI) in animal models but not in clinical trials.We describe a case that the allograft renal vein was twisted because of the surgeon's mistake, which caused the warm ischemia of allograft after reperfusion. The allograft restored blood flow without second reperfusion and cold preservation after 9 min of warm ischemia. The patient was followed up for 3 months and the allograft worked well without complications.

Effects of buried penis on the structure and function of corpus cavernosum in a rat model.

BACKGROUND: While the abnormal appearance of the concealed penis has been well recognized, the effect of buried penis on the structure and function of corpus cavernosum has not been well studied. To explore this issue, we established a rat model and evaluated the effect of buried penis on cavernosum weight, contents and ultrastructure of tissue, and nitric oxide synthase (NOS) activity. METHODS: Two hundred and ten rats were randomly divided into 3 equal cohorts for 2, 4 and 6 months study (groups A, B and C). Each group was randomly divided into buried group (n = 40), control group (n = 15), and normal group (n = 15), respectively. Intra-purse-string suture of the root of the penis was used to establish the model. Macroscopic development was judged by measuring the weight of the corpus cavernosum. Masson's trichrome staining was performed for observing microstructure while a transmission electron microscope was used for observing ultrastructure. The NOS activity was detected by a NOS activity assay kit. RESULTS: Buried penis had no significant influence on the appearance and weight of the corpus cavernosum. Buried penis resulted in decreased smooth muscle content (P > 0.05 in group A, and P < 0.05 in groups B and C) and increased fibrous connective tissue content (P > 0.05 in groups A and B, and P < 0.05 in group C) compared with the normal and control groups. Ultrastructural abnormalities of corpus cavernosum were observed in the 6-month buried group. Moreover, there was decrease of NOS activity in groups B and C (P < 0.05 in group B and P < 0.01 in group C) when compared with the normal and control groups. CONCLUSION: Buried penis affects the structure and function of corpus cavernosum in rats and the effect is positively correlated with the buried time, but there is no significant effect on the macroscopic development.

[Buried penis decreases nitric oxide synthase of the corpus cavernosum].

OBJECTIVE: To explore the influence of buried penis on nitric oxide synthase (NOS) activity of the corpus cavernosum in rats. METHODS: The experimental model of concealed penis was established by intra-pocket-suture of the root of the penis. Two hundred and forty rats were equally randomized into a 2, a 4 and a 6 months group, each further divided into a buried (n = 50), a sham operation (n = 15) and a normal subgroup (n = 15). The development of the corpus cavernosum was surveyed by measuring its weight and the ratio to the body weight, followed by determination of NOS activity in the corpus cavernosum by spectrophotometry. RESULTS: No significant differences were found in the corpus cavernosum weight, the body weight and their ratio among the buried, sham operation and normal groups in any experimental stage (P > 0.05). Buried penis decreased NOS activity in the 4- and 6-month groups (P < 0.05 and P < 0.01) compared with the normal group, but effected no significant change in the 2-month group. CONCLUSION: Buried penis decreases the NOS activity of the corpus cavernosum in a positively time-related manner, but with no significant influence on its appearance and weight.

[Two ways of constructing concealed penis model in rats: a comparative and analytical study].

OBJECTIVE: To establish a stable rat experimental model of concealed penis for studying the effect of buried penis on the structure and function of the corpus cavernosum. METHODS: Ninety male SD rats, aged 2 weeks, were randomly divided into 3 groups (A, B and C) of equal number. Groups A and B underwent surgery with intra-purse suture of the penile root and folding suture of the prepuce, respectively, to bury the penis, while Group C were included as sham operation controls. RESULTS: In Group A, death resulted in 4 cases from acute post-operative urine retention, failure in burying the penis occurred in 5 cases because of soft tissue ulceration around the urethral orifice and in another 3 due to loose concealment. In Group B, 1 died from deep anesthesia and 2 from acute post-operative urine retention. With the penile development and erection, 7 in Group A and 10 in Group B protruded the penis in different stages. In Group C, 1 died from deep anesthesia. The operations succeeded in all the other rats in Groups A and B, with the success rates of 36.7% and 56.7%, respectively. And the concealment could be relieved any time during the experiment. CONCLUSION: The experimental rat model of concealed penis can be successfully established by both intra-purse suture of the penile root and folding suture of the prepuce, which is stable and similar to the natural course of this disorder in human.

[Concealed penis and its surgical correction: a report of 63 cases].

OBJECTIVE: To evaluate the basic principles of surgical correction of concealed penis. METHODS: From Jan. 1999 to Dec. 2002, 63 cases of concealed penis, aged from 1 year and 6 months to 19 years with a mean age of 7 years and 2 months, were corrected with two different approaches: 37 cases of group A with obesity were corrected by anchoring micrus tissue onto the pubis at the base of the penis, and 26 cases of group B by fixing the subcutateous tissues at both sides of the penile shaft to the deep tunica albuginea at the penile root. Then prepuce plasty was carried out. RESULTS: Six-month postoperative follow-up found adequate exposure of the penis in all of group A, prepuce oedema in 7 cases, abdomen fat liquidization in 3 cases, unsatisfactory recovery of incision hollow in 3 cases and patchy numbness of the glans in 1 case. The 25 cases of group B had satisfactory exposure of the penis, with no obvious prepuce oedema, 1 case with retraction of the penis 7 days after operation. CONCLUSIONS: Various surgical procedures could be adopted for concealed penis, and the key to the satisfactory appearance is the restoration of penile tunicae and the fixation at the base of the penis. Further study is needed to determine the natural history of these disorders as well as which conditions and what age will benefit most from surgical intervention.