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

Objective To investigate the therapeutic effects of presurgical TMT on the heights and levels of inferior vena cava(IVC)thrombi,and to assess its impact on surgical strategy.Methods We retrospectively reviewed data of 18 patients with renal cell carcinoma(RCC)involving IVC tumor thrombi who were treated at our hospital with presurgical TMT followed by an IVC thrombectomy.Data from 18 patients(16 men and 2 women)were included in the analysis.The median age was 53.5 years(range:33-75 years),and the mean BMI was 24.7kg/m2(rrange:18.1 -30.4 kg/m2).4 cases of tumors located in the left kidney,14 cases were right.The changes in heights and levels of the IVC thrombi were compared using computed tomography or magnetic resonance imaging.The IVC tumor thrombus level was evaluated according to the Mayo classification.Results The tumor thrombus levels before TMT were stage Ⅰ in 1 patient,Ⅱ in 1 2 patients,Ⅲ in 4 patients,and Ⅳ in 1 patient.The presurgical TMT was sorafenib in 6 patients(33.3%),sunitinib in 9(50.0%),and axitinib in 3(16.7%).After a median of 2 treatment cycles(range:1-6 cycles),three patients experienced grade 3 adverse events.One patient stopped treatment after 6 weeks owing to intolerable skin reactions and difficulty walking.The tumor thrombus height decreased measurably in 11 patients(61.1%).The thrombus height remained stable in 5 patients(27.8%)and was enlarged in 2(11.1%).The median reduction of tumor thrombus height was -0.53 cm (range:-4.23 to 1.21 cm).The median change in the maximum diameter of the thrombus was -0.30 cm (range:-1.23 to 0.29 cm).Down-staging of the thrombus level occurred in 4 patients(22.2%);the surgical strategy was modified in 3 patients(level≥Ⅲ)to avoid cardiopulmonary bypass and complicated liver mobilization under robot-assisted laparoscopy.Conclusions Our data suggest a limited influence of presurgical TMT,with a positive benefit in RCC patients with level Ⅲ and Ⅳ thrombus.Thrombus-level regression may potentially alter the surgical strategy,especially robotic surgery.Additionally,preoperative targeted therapy did not significantly increase perioperative mortality and risk of serious complications.

Objective:To evaluate the efficacy and side effects of PD-1 monoclonal antibody in the treatment of advanced metastatic renal cell carcinoma in China.Methods:The clinical data of 117 patients with advanced metastatic renal cell carcinoma (mRCC) treated with PD-1 monoclonal antibody from October 2016 to February 2022 were retrospectively analyzed. There were 87 males (74.4%) and 30 females (25.6%), with an average age of (57.9±10.9) years old, BMI of (23.6±3.4) kg/m 2and smoking history of 79 (67.5%). There were 44 cases (37.6%) with hypertension, 19 (16.2%) cases of diabetes. The ECOG score of 59.8% (70/117) patients was 0, 33.3% (39/117) was 1, 4.3% (5/117) was 2, and 2.5% (3/117) was 3. The pathological type of 104 cases were renal clear cell carcinoma (ccRCC), 8 cases of papillary renal cell carcinoma, 2 cases of chromophobe cell carcinoma, 2 cases of collecting duct carcinoma and 1 case of eosinophilic cell carcinoma. The general condition of the overall population and the overall survival (OS) of relevant subgroups were analyzed. Secondary goals included progression free survival (PFS), objective response rate (ORR), adverse reactions, overall survival (OS), and progression free survival (PFS). Results:65.8% (77 / 117) of the patients chose targeted combined with PD-1 monoclonal antibody in the first-line treatment. The main targeted drugs were acitinib (81.8%, 63 / 77), tirelizumab (37.6%, 29 / 77) and cindilimab (25.9%, 20 / 77). After first-line treatment, 19.6.1% (23 / 117) patients needed to be converted to second-line treatment, and 15 patients changed the type of PD-1 antibody during treatment. In addition, the targeted drug of combined therapy was replaced by acitinib in 8 patients. The main causes of drug withdrawal were disease progression (70.7%, 29 / 41) and death (29.2%, 12 / 41). The median OS of the overall population was 35.6 (19-60) months and PFS was 12.1 (1-60) months. The ORR of the overall population was 47.8% (56 / 117). 4.2% (5/117) patients had complete remission, another 17.0% (20/117) patients were in stable condition, and 43.5% (51 / 117) patients were in partial remission. In the first-line treatment, the median PFS time of targeted combined with PD-1 monoclonal antibody was 12.6 (1-30) months, the median PFS time of PD-1 single drug immunotherapy was 10.5 (1-60) months. In the second-line treatment, the PFS of patients treated with PD-1 monoclonal antibody was 10.1 (4-19) months, and that of patients treated with PD-1 monoclonal antibody combined with targeted therapy was 11.7 (1-25) months. The most common adverse reactions were elevated blood pressure (18.5%, 23 / 124), followed by hypothyroidism (15.3%%, 19/124), rash (14.5%, 18 / 124), elevated transaminase (10.5%, 13 / 124) and bone marrow suppression (9.7%, 12/124). 9.4% (11 / 117) patients needed to reduce the related adverse reactions by interrupting the treatment control of PD-1 monoclonal antibody.Conclusions:The safety and efficacy of PD-1 monoclonal antibody in domestic patients are better, and the side effects are less. The efficacy and safety of PD-1 monoclonal antibody combined with targeted therapy in the real world population are consistent with many key clinical trials abroad. PD-1 monoclonal antibody combined with targeted drugs can be popularized in the domestic MRCC population.