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Archive > Volume 30 Issue 7 > 2023,30(7):577-585. DOI:10.3872/j.issn.1007-385X.2023.07.005 Prev Next
Basic Research
DJ-1 over-expression promotes proliferation, migration, invasion and epithelial-mesenchymal transformation of human gastric cancer MGC803 cells through PTEN/Akt pathway
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Abstract:
Objective: To investigate the effects of DJ-1 gene over-expression on proliferation, migration, invasion and epithelial-mesenchymal transformation (EMT) of human gastric cancer MGC803 cells and the underlying mechanism. Methods: MGC803 cells with DJ-1 over-expression were constructed by gene transfection technology. Three groups of cells, namely MGC803 group, empty vector group, and DJ-1 over-expression group were set. The effects of DJ-1 gene over-expression on proliferation, clone formation, migration and invasion of MGC803 cells were detected by MTT, plate cloning assay, cell scratch assay and Transwell invasion assay, respectively. The effects of DJ-1 over-expression on the expression levels of DJ-1, PTEN, Akt, p-Akt, Snail, vimentin, E-cadherin, MMP-9 and TIMP-3 were detected by qPCR and Western blot. The morphological changes of MGC803 cells were observed by phase contrast microscope. The effect of DJ-1 over-expression on the growth of MGC803 cell transplanted tumor in vivo was detected in nude mice. Results: MGC803 cells with stable DJ-1 over-expression were constructed successfully. The proliferation ability and number of clones in DJ-1 over-expression group were significantly increased compared with those in MGC803 cell group and empty vector group (all P<0.05); the cell migration distance was significantly increased while the scratch distance was significantly shortened in DJ-1 over-expression group compared with those in MGC803 cell group and empty vector group (all P<0.05); and the migrated and invaded cells of DJ-1 over-expression group were significantly more than those of MGC803 group and empty vector group (all P<0.05). Moreover, the expression of DJ-1 was significantly up-regulated while the expression of PTEN was significantly down-regulated at both the mRNA and protein levels in the DJ-1 over-expression group compared with those in MGC803 group and empty vector group (both P<0.05). There was no significant difference in total Akt protein among all groups (P>0.05), but the expression of p-Akt protein in DJ-1 over-expression group was significantly up-regulated compared with MGC803 group and empty vector group (all P<0.05). In addition, Snail, vimentin and MMP-9 were up-regulated in DJ-1 over-expression group, while E-cadherin and TIMP-3 were down-regulated (all P<0.05). Phase contrast microscopy showed that the number of long spindle cells increased while the number of round and oval cells decreased, and the atypia was more obvious in the DJ-1 over-expression group. In vivo experiments showed that the growth rate of transplanted tumor in DJ-1 over-expression group was significantly accelerated, and the weight of transplanted tumor was significantly increased (both P<0.05) as compared with MGC803 group. Conclusion: DJ-1 over-expression can inhibit the proliferation, migration, invasion and EMT of MGC803 cells both in vitro and in vivo through PTEN/Akt pathway.
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