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Aim To investigate the effect of nifedipine on the formation of autophagosomes in hepatoma cell line Huh-7 and its mechanism.Methods Different concentrations of nifedipine were used to interfere with the proliferation of Huh-7 cells in vitro.The effect of nifedipine on the proliferation of Huh-7 cells was detected by cell proliferation experiment and colony formation experiment.The expressions of Beclin1 and LC3B-Ⅱ were detected by Western blot.The effect of nifedipine on the formation of autophagosomes in Huh-7 cells was observed by laser scanning confocal microscopy.Results Nifedipine significantly inhibited the proliferation of Huh-7 cells in a time-and concentration-dependent manner.The IC50 of nifedipine on day 2 was 22.7 mg·L-1.Nifedipine at the concentration of 25 mg·L-1 significantly reduced the colony formation rate of Huh-7 cells compared with the control group, and the inhibition rate of colony formation was(95.46±0.45)%.Western blot analysis showed that nifedipine significantly up-regulated the protein expression levels of Beclin1 and LC3B-Ⅱ.The amount of autophagosomes in nifedipine group cells were more than that of control group, which was observed by laser scanning confocal microscopy.Conclusions Nifedipine significantly inhibits the proliferation of Huh-7 cells and promotes the formation of autophagosomes, which may be related to the up-regulation of Beclin1 protein expression by nifedipine.
RESUMO
<p><b>OBJECTIVE</b>To explore the pattern of lymph node metastasis (LNM) in advanced Siewert type II adenocarcinoma of the gastroesophageal junction (AGEJ) in order to properly guide lymphadenectomy.</p><p><b>METHODS</b>From January 2009 to Jun 2011, a total of 86 patients with advanced Siewert type II AGEJ underwent radical esophagogastrectomy by thoracic-abdominal incision and two-field lymphadenectomy. The clinical characteristics, pathologic features, LNM, and influence factor of thoracic metastasis were retrospectively analyzed.</p><p><b>RESULTS</b>LNM was observed in 65 of the 86 cases (75.6%). Simple abdominal lymph node metastasis was observed in 49 of the 86 cases (57.0%), simple thoracic lymph node metastasis was 2.3%, and thoracic-abdominal metastasis was 16.3%, with a significant difference between the abdominal and thoracic metastatic patterns. LNM frequency was found in the lymph node groups No. 3, No. 1, No. 7, No. 110, No. 2 and No. 9 (from the highest to the lowest). The incidences of those lymph node metastases were 46.5%, 41.9%, 17.4%, 14.0%, 10.5%, and 5.8%, respectively. Vascular tumor embolus was the only independent factor for thoracic lymph node metastasis.</p><p><b>CONCLUSIONS</b>Abdominal lymph node metastases of advanced Siewert type II AGEJ mainly occur around the proximal stomach and the coeliac trunk. The metastatic rate of distal stomach and splenic perihilar lymph nodes is low, but metastatic rate of the group No.110 lymph nodes is high. The thoracic lymph node metastasis is only correlated with vascular tumor embolus.</p>
