Effect of rapamycin treatment in human seminoma TCam-2 cells through inhibition of G1-S transition.

Mammalian target of rapamycin (mTOR) is an important serine/threonine kinase that plays a critical role in several processes including cell cycle, protein synthesis, and energy metabolism. Due to its multiple roles and general dysregulation in cancer, the mTOR pathway is an important target in cancer therapy. However, studies on mTOR activity in seminoma are limited. Therefore, our aim was to investigate the expression of mTOR signaling pathway proteins in the TCam-2 cell line after rapamycin treatment. TCam-2 cells were treated with different concentrations of rapamycin (control (no rapamycin treatment), 4 nM, 20 nM, 100 nM, 500 nM, and 1000 nM rapamycin) for 48 h and 72 h. mTOR, p-mTOR, P70S6K, p-P70S6K, proliferating cell nuclear antigen (PCNA), and caspase-3 expression levels were analyzed by western blot. Apotosis and cell cycle were analyzed by flow cytometry. After 48 h of rapamycin administration, mTOR activity was significantly decreased at 1000 nM (p < 0.05). In addition, P70S6K acitivity significantly decreased in groups at all rapamycin concentrations (***p < 0.001, ****p < 0.0001). After 72 h of rapamycin administration, mTOR pathway activity were significantly decreased at 100, 500, and 1000 nM rapamycin-treated groups (p < 0.05). Moreover, P70S6K expression decreased in all treatment groups (****p < 0.0001). Caspase-3 expression were similar in all groups. While PCNA expression tended to decrease at 48 h in a dose-dependent manner, this decrease was not significant. We detected decreased PCNA expression at 1000 nM rapamycin at 72 h (p < 0.05). The rate of apoptosis increased especially at 1000 nM rapamycin at 72 h (***p < 0.001). On the other hand, according to the results of the cell cycle experiment, G1 phase arrest was detected at all rapamycin doses at 48 and 72 h (***p < 0.001). Our study indicated that 1000 nM rapamycin may inhibit TCam-2 seminoma cells growth by halting cell proliferation through inhibition of G1-S transition. Therefore, we believe that the findings obtained will contribute to the development of new treatment approaches for seminoma patients in the future and in the process of restoring testicular functions and preserving fertility.

First-line therapy for metastatic colorectal carcinoma: modified FOLFOX4 or FOLFIRI-bevacizumab.

PURPOSE: The aim of this study was to compare the efficacy and toxicity of modified (m) FOLFOX4 (folinic acid, 5-fluorouracil [5-FU], and oxaliplatin) vs. FOLFIRI-B (folinic acid, 5-FU, irinotecan, and bevacizumab) as first-line treatment of metastatic colorectal carcinoma (MCRC). METHODS: The medical records of 89 MCRC patients treated with either mFOLFOX4 (group 1) or FOLFIRI-B (group 2) as first-line chemotherapy were evaluated retrospectively. RESULTS: Complete (CR) plus partial response (PR) were seen in 18 (36.7%) vs. 13 (32.5%) patients in the mFOLFOX4 vs. FOLFIRI-B, respectively (p=0.67). Median progression-free survival (PFS) was 9 months (95% CI 7.2- 9.5) vs. 10 months (95% CI 7.6-12.3) in group 1 vs. group 2, respectively (p=0.30). Median overall survival (OS) was 22 months (95% CI 17.6-26.3) and 19 months (95% CI 13-24.9) in group 1 and 2, respectively (p=0.32). There was no statistically significant difference in grade 3-4 hematological toxicity between the groups, but grade 3-4 grade weakness, diarrhea, nausea and vomiting was observed more frequently in the FOLFIRI-B patients (p=0.03, p=0.01, p=0.05, respectively). CONCLUSION: Our data suggest that mFOLFOX4 and FOLFIRI-B are equally effective as first-line chemotherapy in MCRC patients. This may partially be explained by the fact that almost 50% of those receiving FOLFOX in the first-line received FOLFIRI-B in the second-gline, an observation suggesting that bevacizumab in the second line may be as effective as in the first line.

The effect of different treatment modalities on survival in elderly patients with locally advanced non-small cell lung cancer.

PURPOSE: The aim of this study is to investigate the effect of treatment modalities on survival among unoperat ed and locally-advanced non-small cell lung cancer (NSCLC) patients aged 70 years and older, representing real-life data. METHODS: From 2005 through 2017, medical records of 2259 patients with lung cancer from Okmeydani Training and Research Hospital-Istanbul/Turkey were reviewed retrospectively. Patients with locally advanced NSCLC ≥ 70 years of age who did not undergo surgery for lung cancer were reviewed. In total, 130 patients were eligible for the final analysis. Patients were stratified into four groups as: chemotherapy (CT), concurrent chemoradiotherapy (cCRT), sequential chemoradiotherapy (sCRT), and radiotherapy (RT) only. RESULTS: Of the 130 patients included in the analysis; CT, cCRT, sCRT, and RT only were applied to 25(19.2%), 30(23.1%), 31(23.8%), and 44(33.8%) patients, retrospectively. Twelve (9.2%) patients were female. Median age was 72 years (range, 70-88). Sixty (46.2%) patients had stage IIIA disease and 70(53.8%) patients had stage IIIB disease. Median progression-free survival(mPFS) in patients treated with CT, cCRT, sCRT, and RT were 8.0, 15, 10, and 9.0 months, respectively(p = 0.07). Corresponding median overall survival (mOS) were 10, 33, 20, and 15 months (p = 0.04). In multivariate analysis, stage IIIB disease [hazard ratio (HR), 2.8], ECOG-PS 2(HR, 2.10), and ECOG-PS 3-4(HR, 5.13) were found to be the negative factors affecting survival, while cCRT (HR, 0.45) and sCRT (HR, 0.50) were the independent factors associated with better survival. CONCLUSION: This study showed that the use of combined treatment modality was associated with better survival in elderly patients with locally advanced NSCLC, with the greatest survival observed in patients treated with cCRT. We therefore suggest that cCRT, when feasible, should be strongly considered in locally advanced NSCLC patients 70 years and over.

The correlation between tumor markers and Ga-68 DOTA-TATE PET/CT findings in patients with advanced medullary thyroid cancer. / La correlación entre los marcadores tumorales y los hallazgos de Ga-68DOTA-TATE PET/TC en pacientes con cáncer medular de tiroides avanzado.

BACKGROUND: Medullary thyroid cancer is a rare type of neuroendocrine tumor. Calcitonin (Ctn) and carcinoembryonic antigen (CEA) are used as markers for medullary thyroid cancer. The aim of the study was to evaluate the importance of serum Ctn and CEA levels in predicting total tumor volume. MATERIAL AND METHODS: Ga-68 DOTA-TATE PET/CT findings such as whole-body somatostatin receptor-expressing metabolic tumor volume (SSR-E MTV) and total lesion volume (SSR-E TLV) were calculated and correlation analysis was done for tumor markers and whole-body SSR-E MTV and TLV. RESULTS: A total of 28 patients with advanced medullary thyroid cancer were included in this retrospective study. In the correlation analysis, there was a statistically significant positive correlation between Ctn and whole-body SSR-E MTV (rho=0.503 and P=.008). Similarly, significant positive correlation was observed between Ctn and whole-body SSR-E TLV (rho=0.436 and P=.023). There was a significant positive correlation between CEA and whole-body SSR-E MTV (rho=0.584 and P=.007). Also, a positive correlation was observed between CEA and whole-body SSR-E TLV (rho=0.436 and P=.023). These correlations were most marked in patients with Ctn≥152pg/mL and/or both nodal and bone involvement. CONCLUSION: These results could stimulate clinicians to perform Ga-68 PET for treatment decisions, especially in patients with high CEA and Ctn levels at the time of diagnosis.

Sirtuin 1 independent effects of resveratrol in INS-1E ß-cells.

Resveratrol (Resv), a natural polyphenol, is suggested to have various health benefits including improved insulin sensitivity. Resv activates Sirtuin (Sirt1) in several species and tissues. Sirt1 is a protein deacetylase with an important role in ageing, metabolism and ß-cell function. In insulinoma ß-cells (INS-1E), Resv is previously shown to improve glucose-stimulated insulin secretion in a Sirt1-dependent mechanism and to protect against ß-cell dedifferentiation in non-human primates, while inducing hypertrophy in myoblasts. Mammalian (mechanistic) Target of Rapamycin (mTOR), is a key regulator of cellular metabolism and regulates the cell size, ß-cell survival and proliferation. In order to understand the interaction of Sirt1 and mTOR cascade activity with Resv-induced changes in the INS-1E cell line, we generated stable Sirt1-down-regulated INS-1E cells, and analysed Sirt1-dependent effects of Resv with respect to mTOR cascade activity. Sirt1-knockdown (KD) had a significant increase in cell size compared to negative-control (NEG CTR) cells. Resveratrol treatment increased cell size in both cell types in a dose-dependent manner at 24 h (Resv conc: 15-60 µM), and decreased the cell number (Resv conc: 30-60 µM). Cell area was increased in NEG CTR cells (Resv conc: 60 µM) at 24 h and KD cells at 48 h (Resv conc: 15-60 µM). Rapamycin, a specific mTOR inhibitor, counteracted the Resv-induced cell enlargement (both cell diameter and area). Furthermore, Sirt1-downregulation by itself did not affect the mTOR cascade activities as measured by Western blotting for total and phosphorylated Akt and mTOR. Rapamycin decreased the mTORC1 activity, while increasing the pAkt levels. Resveratrol did not interfere with the mTOR activity or with Sirt1 expression. Altogether, this work indicates that Sirt1 is a negative regulator of cell size. Moreover, the effect of Resv on cell size increase is mTOR-cascade dependent.

Surgical glove-port single-incision laparoscopic gastric wedge resection of gastrointestinal stromal tumors: initial experience with 2 cases.

Gastrointestinal stromal tumors are localized mainly in the stomach, and the therapeutic approach is surgical resection. Laparoscopy can be performed for tumors located in the greater curvature of the stomach. Among the treatment alternatives, a single-incision laparoscopic technique is used more frequently than standard laparoscopy due to the successful results of laparoscopic surgery. Here, we report the use of single-incision surgical glove-port laparoscopy for the resection of 2 gastrointestinal stromal tumors localized in the greater curvature of stomach.