OBJECTIVETo construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.

METHODSLentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.

RESULTSThe recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).

CONCLUSIONVHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Apoptosis ; Carcinoma, Renal Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

OBJECTIVETo investigate the expression of miR-223 in clear cell renal cell carcinoma (ccRcc) and its clinical implications.

METHODSQuantitative real-time PCR was employed to detect the levels of miR- 223 expression in ccRcc, pair-matched adjacent normal tissues and different renal cancer cell lines. Transwell migration essay and wound healing essay were used to evaluate the invasion and migration of renal cancer 786-O cells transfected with miR-223 mimics. MTT essay was used to measure the cell proliferation, and the cell cycle changes following the transfection were analyzed with flow cytometry.

RESULTSCompared with the normal tissues, the cancer samples showed up-regulated miR-223 expression, which was associated with tumor size. In 786-O cell cultures, transfection with miR-223 mimics significantly enhanced cell migration (P<0.0001) and growth (P=0.006) and induced G1 cell cycle arrest.

CONCLUSIONmiR-223 promotes renal cancer cell migration and proliferation and may serve as a potential therapeutic target for ccRcc.

Carcinoma, Renal Cell ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; G1 Phase Cell Cycle Checkpoints ; Humans ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation

Objective:To evaluate the efficacy of holographic image technology in robot-assisted laparoscopic radical prostatectomy (RARP).Methods:The clinical data of 34 patients with prostate cancer who underwent RARP in our hospital during October 2020 and December 2020 was analyzed retrospectively. The average age of the patients was 67.8 (52-78) years. The mean BMI was 25.8 (18.0-32.3) kg/m 2. The median level of PSA before surgery was 13.4 (2-149) ng/ml. Median prostate volume was 31.7 (9.5-159.1) ml. EAU risk groups for biochemical recurrence of localised and locally advanced prostate cancer were list as below: 5 cases of low-risk, 7 cases of medium-risk, 22 cases of high-risk. There were 9, 16, 9 cases with the ASA score of 1, 2, 3 point, respectively. Preoperative Gleason score of 34 patients were list as below: 9 cases in score ≤6 group, 15 cases in score=7 group, 10 cases in score ≥8 group. For clinical stage before the surgery, 13 cases ≤T 2a stage, 1 case in T 2b stage, 20 cases ≥T 2c stage. The engineers established holographic images of 34 patients based on multiparametric magnetic resonance imaging (mpMRI) and the reports before the operation. Surgeons can obtain the size and location of tumors, surrounding neurovascular bundles visually by revolving, assembling, disassembling and concealing images, which was helpful for pre-surgery planning. By manipulating the holographic images extracorporeally, surgeons can discriminate Internal sphincter of urinary bladder and vesicoprostatic muscle, neurovascular bundles, membranous part, seminal vesicle easily, which improves the operation accuracy. Results:All 34 cases underwent operation successfully without transferring to open surgery. The median operative time was 157.5 (95-276) min with an estimated blood loss of 50 (20-300) ml. The median drainage removal time was 2 d and median hospitalization time was 3.5 d, respectively. The catheters were removed within an average time of 20.5 d. For postoperative Gleason score, there were 2 cases in score ≤6 group, 16 cases in score =7 group, 8 cases in score ≥8 group and 8 cases can’t make a score. For clinical stage after the surgery, 10 cases were ≤T 2a stage, 1 case was T 2b stage, 23 cases were ≥cT 2c stage. 22 cases underwent pelvic lymph node dissection, including a patient with right iliac fossa lymph node metastasis. There were 2 cases with positive surgical margin and 3 cases with Clavien-DindoⅠcomplications. The rate of 1-month and 3-month urinary continence were 47.1% and79.4%, respectively, 8 cases recovered erectile function after 3 month. Conclusions:Holographic image technology can promote cancer dissection completely, achieve urinary continence early and reduce perioperative complications tremendously. The technology is the "intraoperative security" for the accurate surgical treatment of prostate cancer.

Objective:To investigate the characteristics of 18F-DCFPyL PET/CT imaging in castration-resistant prostate cancer (CRPC) patients with different PSA levels. Methods:The imaging and clinical data of 50 patients with CRPC who underwent 18F-DCFPyL PET/CT examination in Chinese PLA General Hospital from January 2018 to December 2020 were analyzed retrospectively. The average age was 72 (54-95) years old. Serum total PSA was 92.28(0.36-2000.00) ng/ml. According to the total PSA level, the patients were divided into low PSA group(total PSA ≤ 1 ng/ml, n=9), medium PSA group (1 ng/ml10ng/ml, n=23). According to the standardized evaluation standard of molecular imaging, the suspicious tumor lesions on 18F-DCFPyL PET/CT imaging were scored by molecular imaging PSMA(miPSMA), and the miPSMA score ≥2 was defined as positive lesions. According to the number of lesions displayed by 18F-DCFPyL PET/CT, patients were divided into oligofocal group (the number of lesions ≤3) and multiple lesions group (the number of lesions >3). The imaging characteristics of patients in different groups were summarized. Results:The 18F-DCFPyL PET/CT imaging results of 50 cases in this study were all positive, including oligofocal group (n=27) and multiple lesions group (n=23). Of the 30 patients with unresected prostate, 18 had local recurrence of the prostate, while the other 12 patients with unresected prostate and 20 patients with resected prostate had no signs of local recurrence. The oligofocal group showed local recurrence, regional lymph node metastasis or bone metastasis. Patients with multiple lesions showed multiple lymph nodes and/or bone metastasis with or without local recurrence. There were 9, 18 and 4 patients with oligofoci in low, middle and high PSA groups, respectively.There were 27 patients in the oligonucleogenous group, and 21 of the 22 patients receiving local treatment were effective. All 3 patients treated with systemic treatments were effective. PSA progressed in 2 untreated patients. In the multi-foci group of 23 patients, 6 of 9 patients treated with abiraterone were effective. Two patients treated with enzalumide were ineffective. One of the 4 patients with chemotherapy was effective. One of the two patients treated with 177 Lu-PSMA nuclide was effective. One case did not respond to treatment with 89SrCl 2. Radiotherapy failed in 2 cases. PSA progressed in 3 untreated patients. Conclusions:18F-DCFPyLPET/CT imaging has a high detection rate of lesions in patients with CRPC and has potential guiding significance for follow-up treatment. The number of lesions in CRPC patients with different PSA levels was different, and the patients with low PSA levels were mainly oligofoci.

【Objective】 To statistically analyze the relationship between homologous recombination repair deficiency (HRD) score and clinicopathological characteristics, genomic mutations in patients with high-risk and metastatic hormone-sensitive prostate cancer (mHSPC) and the prognostic predictive value in mHSPC. 【Methods】 A total of 127 patients diagnosed with high-risk prostate cancer and mHSPC, treated at the Department of Urology of Chinese PLA General Hospital during Dec.2021 and Nov.2023 were enrolled.Homologous recombination repair (HRR) gene sequencing was performed, and the genomic scar score (GSS) algorithm were conducted to calculate the HRD score.The relationship between HRD scores and clinicopathological features, genomic alterations, and prognosis were analyzed. 【Results】 The median HRD score was 1.6(0.8, 5.2), 30(23.6%) patients’ HRD scores ≥10, and 11(8.7%) patients’ HRD scores ≥20.Clinicopathological features, including ISUP classification ≥4 (P=0.044) and metastatic status (P=0.008) were associated with high HRD score.Patients with mutations in the BRCA, TP53 and MYC systems had significantly higher HRD score than those with wild-type genes (P<0.05).In mHSPC, the risk of biochemical recurrence was 12.836 times higher in patients with HRD score ≥20 than in those with <20 [OR:12.836 (1.332-124.623), P=0.028]. 【Conclusion】 Baseline HRD score was lower in patients with high-risk prostate cancer and mHSPC.Patients with high HRD score may have higher histological grading (ISUP≥4) and later clinical stage.Further investigation is needed to determine the threshold of HRD scores as biochemical markers suggestive of a poor prognosis.