| ObjectiveGastric cancer is one of the most common primary gastrointestinal cancers in the world. Recurrenceand metastasis are the basic reasons leading to pool prognosis of advanced gastric cancer. In recent years,epithelial-mesenchymal transition (EMT) has been found to be closely associated with tumor metastasis,and also been regarded as an important mechanism of tumor metastases. Nowadays, many genesmodulating gastric cancer metastasis have been reported, yet the mechanism of tumor metastases remainspoorly understood as a result of its complexity. Thus, to find new aberrant genes involved in the regulationof gastric cancer metastasis is very significant for diagnosis, treatment and prevention against themetastasis and recurrence of gastric cancer. Scinderin is a Ca2+-dependent filamentous actin (F-actin)severing and capping protein, which plays a key role in secretion. But few researches are about biologicaleffects of scinderin on human disease especially neoplasm. This study will reveal the effects of silencingscinderin on the migration, EMT process, proliferation and cell cycle distribution of highly metastatichuman gastric cancer cell line SGC-7901.Methods1. To establish the stable scinderin-silencing SGC-7901cells, the shRNA targeting scindern lentiviralvector was constructed, and then transfected into highly metastatic human gastric cancer cell lineSGC-7901. The efficiency of transfection was observed under fluorescence microscope. Effects of genesilencing were confirmed by RT-qPCR and Western blot.2. The effect of scinderin knockdown on migration in SGC-7901cells was analyzed by Transwellassay.3. After scinderin knockdown in SGC-7901cells, the changes in the expression of EMT markers (suchas E-cadherin and N-cadherin) were investigated by RT-qPCR and Western blot.4. The effect of scinderin knockdown on proliferation in SGC-7901cells was analyzed by real-timecell analyzer (RTCA), and the cell cycle distribution was investigated by flow cytometer.Results1. Scinderin-shRNA lentiviral vector was successfully transfected into gastric cancer cell lineSGC-7901, and levels of scinderin mRNA and protein expression reduced significantly (P<0.01). Theseresults suggested that the stable scinderin-silencing gastric cancer cell line SGC-7901had beensuccessfully established.2. Transwell migration assay indicated that scinderin knockdown could attenuate the migration ofSGC-7901cells (P<0.05).3. RT-qPCR and Western blot assay indicated that the expression of E-cadherin in scinderin-silencingSGC-7901cells was significantly up-regulated (P<0.001), while N-cadherin expression obviouslydecreased (P<0.001). Furthermore, scinderin knockdown in SGC-7901cells could cause the lowerexpression of β-catenin protein (P<0.01).4. RTCA proliferation assay indicated that scinderin knockdown could significantly suppress the proliferation of SGC-7901cells (P<0.05). And cell cycle analysis by flow cytometer suggested thatscinderin knockdown in SGC-7901cells was able to arrest cell cycle at G2/M phase.Conclusions1. Silencing scinderin can attenuate EMT process of highly metastatic human gastric cancer cell lineSGC-7901, and effectively restrain their migration.2. Silencing scinderin can arrest the cell cycle of SGC-7901cells, and significantly reduce theirproliferation.3. This study suggests that scinderin is closely related to the metastasis of gastric cancer cells and theregulation of tumor EMT process, which contributes to illuminating the mechanism of gastric cancermetastasis and becoming a very important practical tool for diagnosis, treatment and prevention against themetastasis and recurrence of gastric cancer someday. |
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