Abstract
Gastric cancer metastasis remains a major cause of cancer-related deaths. There is an urgent need to develop new therapeutic approaches targeting metastatic gastric cancer. Argininosuccinate synthetase 1 (ASS1) expression is increased in gastric cancer. We detected the protein expression of ASS1 in human gastric cancer cell lines (AGS, NCI-N87, and MKN45) and in murine gastric cancer cell lines (3I and 3IB2). We used vector-mediated short hairpin RNA (shRNA) expression to silence ASS1 expression in the MKN45 and 3IB2 cell lines, and analyzed the effects of this protein on cell migration and metastasis. We demonstrated that ASS1 silencing suppressed cell migration in the MKN45 and 3IB2 cell lines. ASS1 knockdown significantly reduced liver metastasis in mice after the intrasplenic implantation of 3IB2 cancer cell clones. To determine whether arginine restriction may represent a therapeutic approach to treat gastric cancer, the sensitivity of tumor cells to arginine depletion was determined in gastric cancer cells. Arginine depletion significantly inhibited cell migration in the gastric cancer cell line. The silencing of ASS1 expression in MKN45 and 3IB2 gastric cancer cells markedly decreased STAT3 protein expression. In conclusion, our results indicate that the ASS1 protein is required for cell migration in gastric cancer cell lines.
Highlights
Gastric cancer metastasis remains a major cause of cancer-related deaths
Compared with the vector control and nonsilencing RNA interference (RNAi), the Argininosuccinate synthetase 1 (ASS1) RNAi-1 and ASS1 RNAi-2 decreased the protein levels of ASS1 (Supplementary Figure S1b). These data suggest that lentivirus-mediated ASS1 RNAi effectively inhibits endogenous ASS1 expression in MKN45 cells
ASS1 silencing led to a decrease in the expression of the ASS1 protein in MKN45 cancer cells compared with parental and vector control (VC) cells (Figure 1a and Supplementary Figure S2)
Summary
Gastric cancer metastasis remains a major cause of cancer-related deaths. There is an urgent need to develop new therapeutic approaches targeting metastatic gastric cancer. We detected the protein expression of ASS1 in human gastric cancer cell lines (AGS, NCI-N87, and MKN45) and in murine gastric cancer cell lines (3I and 3IB2). We used vector-mediated short hairpin RNA (shRNA) expression to silence ASS1 expression in the MKN45 and 3IB2 cell lines, and analyzed the effects of this protein on cell migration and metastasis. We determined ASS1 expression in three different human gastric cancer cell lines (AGS, NCI-N87, and MKN45) and in a murine gastric cancer cell line (3IB2) that was originally derived from an orthotopic transplantable gastric cancer in ICR mice[9,10]. We used an RNA interference (RNAi) approach to target ASS1, a key enzyme involved in arginine metabolism, in the MKN45 and 3IB2 cell lines. Experiments using a 3IB2 murine gastric cancer model further supported the notion that ASS1 is essential for metastasis
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