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【Objective】 To investigate the effects of the loss of exon 1 of TFE3 on nuclear localization of chimeric TFE3 protein in TFE3 translocation renal cell carcinoma (TFE3 tRCC). 【Methods】 The localization of TFE3 protein in TFE3 tRCC and clear cell renal cell carcinoma (ccRCC) were detected with immunochemistry. The exon retention of TFE3 gene in TFE3 tRCC was analyzed in databases and literatures. The plasmids containing TFE3 full-length and different-length of TFE3 exons which were constructed to pCDH-MCS-EGFP-Puro were transfected into HEK293T using Lipo FiterTM. The localization of EGFP protein in HEK293T cells were detected with confocal microscopy. The localization of TFE3 protein and truncated TFE3 protein were detected with Western blotting. The mRNA expression of the downstream genes of TFE3 protein were detected with q-PCR. 【Results】 Strong nuclear signal of TFE3 protein was observed in TFE3 tRCC, whereas TFE3 protein in ccRCC was mainly localized in cytoplasm. The results of fluorescence imaging and Western blotting showed that TFE3 full-length protein was expressed both in nucleus and cytoplasm, and the expression of truncated TFE3 protein was mainly localized in nucleus. The q-PCR analysis demonstrated that the deletion of exon 1 in TFE3 gene led to a higher transcriptional level of targeted genes of TFE3 protein. 【Conclusion】 The loss of exon 1 in TFE3 played a critical role in preventing TFE3 protein from entering the nucleus. In TFE3 tRCC, the loss of exon 1 in TFE3 gene leads to the nuclear localization of TFE3 fusion protein and activation of its downstream target genes. This mechanism promises to uncover the occurrence and development of TFE3 tRCC.
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Objective:A single-center analysis was performed to assess the significant clinical features and prognostic factors of TFE3-rearranged renal cell carcinoma (TFE3 rRCC).Methods:The clinical data of 85 confirmed cases of TFE3 rRCC patients at the Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University from January 2007 to February 2023 were analyzed retrospectively. Among these patients, there were 39 males and 46 females, with a median age of 32 (26, 45) years. All patients underwent preoperative CT scans, 21/85 cases (24.7%) of TFE3 rRCC exhibited the characteristic feature of "circular calcification" with plain CT imaging, and enhanced CT scan showed that the tumor enhancement during the arterial phase was lower than the adjacent renal cortex. Among the 85 patients in this cohort, the median tumor diameter was 4.8(3.2, 6.5). Thirty-two patients underwent partial nephrectomy (NSS), while 51 patients underwent radical nephrectomy (RN). Two patients with distant metastasis at the time of diagnosis received only sunitinib therapy. Forty-three patients received adjuvant treatment, including 14 patients who received targeted therapy. There were 29 patients in AJCC stage Ⅲ/Ⅳ, with 10 patients presenting with venous tumor thrombus and 14 patients with lymph node metastasis. Histopathology, TFE3 immunohistochemistry, and break-apart TFE3 FISH probe detection were performed on all 85 cases, while 52 patients underwent RT-PCR and/or DNA sequencing. By combining the clinical and pathological data, we summarized the diagnostic Methods for TFE3 rRCC, evaluated the impact of surgical approaches (RN and NSS) on the survival outcomes of cT 1a/b patients, and assessed the influence of genetic subtypes (ASPL, NONO, PRCC, SFPQ, and others) on the survival outcomes of all patients. Furthermore, we analyzed the risk factors for disease progression. Results:TFE3 rRCC exhibited variable histopathological features, and the presence of acinar-like structures with psammoma bodies may be a relatively typical characteristic. All 85 patients showed positive TFE3 immunohistochemical staining. In 6 cases of TFE3 rRCC, break-apart TFE3 FISH probe yielded negative results. However, final confirmation was achieved through genetic sequence, with 5 cases diagnosed as NONO-TFE3 subtype and 1 case as RBM10-TFE3 subtype. Among the 85 patients, 52 underwent RT-PCR and/or DNA sequencing, revealing a total of 8 TFE3 fusion subtypes, including 11 cases of ASPL-TFE3, 8 cases of PRCC-TFE3, 10 cases of NONO-TFE3, 15 cases of SFPQ-TFE3, 1 case of CLTC-TFE3, 2 cases of LUC7L3-TFE3, 4 cases of MED15-TFE3, and 1 case of RBM10-TFE3. The survival analysis results revealed that among the 12 patients with cT 1b stage tumors who underwent radical nephrectomy (RN), the progression-free survival (PFS) was 35 (14, 109) months, which was significantly better than the NSS group ( P=0.041). However, for the 14 patients with cT 1a stage tumors who underwent RN, there was no statistically significant difference in overall survival (OS) and PFS compared to the NSS group, with OS being 55(27, 134) months and PFS being 71(41, 134) months. Stratifying according to TFE3 fusion subtypes, it was found that patients with ASPL-TFE3 fusion had a significantly lower PFS compared to those with non-ASPL-TFE3 fusion subtypes ( P=0.029). Survival analysis revealed that tumor diameter, surgical approach, adjuvant therapy, AJCC staging, venous tumor thrombus, and lymph node metastasis were associated with OS and PFS( P<0.05). The results of the multivariate Cox regression analysis showed that AJCC stage Ⅲ/Ⅳ( HR=2.393, 95% CI 1.418-4.039, P=0.001) and venous tumor thrombus ( HR=3.543, 95% CI 1.159-10.827, P=0.026) were independent risk factors for progression-free survival (PFS). Conclusions:During the non-enhanced phase of CT scan, TFE3 rRCC tumors can exhibit a circular calcification. TFE3 immunohistochemistry is an important screening method for TFE3 rRCC.Break-apart TFE3 FISH probe detection is considered the diagnostic gold standard, and gene sequencing, if feasible, can provide the subtype diagnosis of TFE3 rRCC. For cT 1a stage TFE3 rRCC, partial nephrectomy (NSS) is a viable option, while caution should be taken in selecting NSS for cT1b stage patients. Patients with ASPL-TFE3 fusion subtype have a worse prognosis. AJCC stage Ⅲ/Ⅳ and venous tumor thrombus indicate poor prognosis in TFE3 rRCC.
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Objective:The prognosis of cancer patients depends not only on tumor related factors, but also on host related factors, especially systemic inflammatory response. Based on the ratio of neutrophils to lymphocytes (NLR), the ratio of platelets to lymphocytes (PLR) and the ratio of lymphocytes to monocytes (LMR), we constructed a systemic inflammation model to predict the survival time of patients with gastric cancer (GC) after radical gastrectomy.Methods:Two hundred and five patients with GC who underwent radical resection from January 2011 to January 2017 were selected in Qinghai Provincial Communications Hospital and Red Cross Hospital of Qinghai Province. NLR, PLR and LMR were collected before operation. The best truncation values of NLR, PLR and LMR were obtained by ROC curve and systemic inflammatory marker score (SIMS) was constructed. The clinical value of SIMS was analyzed by single factor and multi factor Cox risk proportion model.Results:All patients were followed up for an average of (63.47 ± 10.36) months (range 20 to 65 months). The median survival time was 56 months. The one-year mortality rate was 6.3%, the three-year mortality rate was 26.2%, and the five-year mortality rate was 34.6%. The AUC of NLR, PLR and LMR were 0.745, 0.805 and 0.866 respectively, and the best truncation values were 3.11, 144 and 3.34 respectively. The mortality of patients with NLR > 3.11, PLR > 144, LMR ≤ 3.34 was higher than that of patients with NLR ≤ 3.11, PLR ≤ 114, LMR > 3.34 ( χ2 = 10.491, 14.658 and 38.765; P<0.01); there were differences in survival curves among different groups of NLR, PLR, LMR ( P < 0.05). The survival curves of different scores of SIMS were different ( P < 0.05). Age ( HR = 1.358, 95% CI 1.153 to 1.599), T stage-T 3 ( HR = 2.739, 95% CI 1.200 to 6.248), T stage-T 4 ( HR = 3.013, 95% CI 1.312 to 6.920), N stage-N 2 ( HR = 5.832, 95% CI 2.974 to 11.455), pathological stage Ⅲ ( HR = 2.962, 95% CI 1.835 to 4.646), lymphovascular invasion ( HR = 1.813, 95% CI 1.274 to 3.642), SIMS-1 ( HR = 7.065, 95% CI 4.673 to 10.692), SIMS-2 ( HR = 7.885, 95% CI 4.991 to 12.435), SIMS-3 ( HR = 8.365, 95% CI 5.635 to 3.485) were the independent risk factors of GC patients′ death ( P < 0.05). Conclusions:This study successfully constructs Sims and confirms that preoperative Sims is a relatively easy, easy to obtain and low-cost prognosis index for GC patients, which can be used to evaluate the survival time of GC patients before operation.
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Objective To determine whether nitric oxide may result in different effects on cardiac function at the different time points during the course of myocardial ischhemia and reperfusien of rats in viwo, and if so, to investigate the mechanisms involved.Methods Male adult rats were subjected to 30 minutes of myocardial ischemia followed by 5 hours of reperfusien. L-arginine was administrated at a bolus at both 10 minutes before and 50 minutes after reperfusion in early treatment group. The L-arginine was administrated at a bolus at both 3 and 4 hours after reperfusion in late treatment group. The following variables were measured: cardiac hemodynamic parameters, myocardial easpase 3 activity, iNOS and total NO content. Results Compared with vehicle group, administration of L-arginine at early reperfusion markedly reduced myocardial caspase 3 activity, and improved cardiac function. In strict contrast, administration of L-arginine at late reperfusion resulted in a significant increase in myocardial NOx content, myocardial apoptosis, and depressed the cadiac function. Conclusion Timely administration of L-arginine might inhibit apoptosis while late administration of L-arginine might enhance apoptosis.