SPINK1 contributes to proliferation and clonal formation of HT29 cells through Beclin1 associated enhanced autophagy.

We aimed to explore the molecular mechanism that were involved in SPINK1-induced proliferation and clonogenic survival of human colorectal carcinoma (CRC) HT29 cells. Initially, we generated HT29 cells either permanently silencing or overexpressing SPINK1 protein. The results showed that SPINK1 overexpression (OE) significantly stimulated the proliferation and clonal formation of HT29 cells at the varied time points. Secondly, we found SPINK1 OE enhanced the ratio of LC3II/LC3I and the level of autophagy-related gene 5 (ATG5), whereas SPINK1 knockdown (Kd) reversed the above outcome under normal culturing and/or fasting condition in the cells, indicating its role in autophagy enhancement. Moreover, LC3-GFP-transfected SPINK1-OE HT29 cells strengthened the fluorescence intensity compared with the untransfected control. Chloroquine (CQ) significantly decreased the level of autophagy in both control and SPINK1-OE HT29 cells. The autophagy inhibitors, CQ and 3-Methyladenine (3-MA), remarkably inhibited the proliferation and colony formation of SPINK1-OE HT29 cells, while ATG5 upregulation resulted in the growth of the cells, suggesting the important function of autophagy in cell's growth. Thirdly, SPINK1-induced autophagy was independently of mTOR signaling as p-RPS6 and p-4EBP1 were activated in SPINK1-OE HT29 cells. Instead, Beclin1 up and down regulation were clearly observed in SPINK1-OE and SPINK1 Kd HT29 cells, respectively. Moreover, Beclin1 silencing apparently reduced autophagy in SPINK1-OE HT29 cells, indicating that SPINK1-induced autophagy was closely associated with Beclin1. Collectively, SPINK1-promoted proliferation and clonal formation of HT29 cells were closely associated with Beclin1 associated enhanced autophagy. The above findings would open a new window for probing the role of SPINK1-related autophagic signaling in the pathogenesis of CRC.

Outcomes of local thoracic surgery in patients with stage IV non-small-cell lung cancer: A SEER-based analysis.

BACKGROUND: The outcomes of thoracic surgery for patients with stage IV non-small-cell lung cancer (NSCLC) are controversial and uncertain. PATIENTS AND METHODS: The National Cancer Institute's Surveillance, Epidemiology, and End Results was queried for patients with stage IV NSCLC, including those treated with surgery-participated therapy modalities. Overall survival (OS) was evaluated using a variety of statistical analyses. RESULTS: The analysis was carried out for 90,982 patients from 1975 to 2016 who had been diagnosed as stage IV NSCLC. Propensity score-matched (PSM) analyses that were well-balanced with all the important confounding covariates revealed improved OS (median survival time [MST]) with patients receiving surgery versus non-surgery (MST: 15 versus 8 months, P < 0.001); undergoing surgery plus chemotherapy versus chemotherapy (MST: 19 versus 11 months, P < 0.001); and having surgery plus chemoradiation versus chemoradiation (MST: 18 versus 11 months, P < 0.001). Sequential landmark analyses for long-term survivors of ≥1 and ≥3 years all indicated improved OS (P < 0.001) on univariate and multivariate analyses for the patients receiving the three surgery-related treatment patterns listed earlier, relative to the corresponding surgery-absent treatment modalities. For synchronous presentations of varied treatment paradigms, surgical intervention significantly led to increased OS (MST, months) benefits following treatment paradigms: surgery plus chemotherapy (22), surgery plus chemoradiation (18), chemotherapy (10), surgery only (9), chemoradiation (9), surgery plus radiation (6) and radiation alone (2). The subgroup analysis demonstrated that the elevated OS associated with local thoracic surgery in addition to chemotherapy (versus chemotherapy) or chemoradiation (versus chemoradiation) fell in the subcategories of T0-3, N0-2 and 0-1 (metastatic sites) tumours. The comparison of the aforementioned two types of treatment patterns indicated that the optimal patients for the surgery were those with any combination of T1-4, N0-3, Msite0-1 and adeno- or squamous carcinoma. CONCLUSIONS: The patients with T1-4, N0-3, Msite0-1 and adeno- or squamous carcinoma of stage IV NSCLC had a longer OS with local thoracic surgery in combination with chemotherapy or chemoradiation.