| ObjectiveGastric cancer(GC) is a common malignant digestive cancer and the third leading causes of cancer related death worldwide. Although the incidence of gastric cancer now have fallen, but the prognosis of patients with gastric cancer is still not ideal. Because of the gastric cancer metastasis and recurrence risk is higher and the lack of effective biomarker detection of early gastric cancer, the mortality of gastric cancer is still high. Modern biomedical research has explored many potential gastric cancer biomarker genes by utilising serum protein antigens, oncogenic genes or gene families through improving molecular biological technologies, such as microarray, RNA-Seq and the like. Vimentin plays important roles in the epithelial to mesenchymal transition(EMT) and cancer growth. This research mainly study vimentin, USP14 and miR-320 a in gastric cancer cells in malignant behavior, explore their mutual adjust network structure in gastric cancer, we hope through our study can reasrch for the early diagnosis and treatment of gastric cancer and put forward a new train of thought. MethodFirstly, application RT-qPCR detection of vimentin expression in gastric cancer tissue level, and studied the role of vimentin in gastric malignant behavior. Then, we explored the mechanism of vimentin expression differences between gastric cancer tissues and normal gastric tissue, we used ubiquitin and Co-immunoprecipitation assays to explore the mechanism of vimentin discrepant expression level, finding USP14 through influencing the ubiquitin of vimentin to affect it level in gastric cancer. Moreover, the MTT, colony forming experiment, migration and invasion experiments respectively use to determine that USP14 facilitated cell proliferation, migration and invasion ability in the gastric cancer cell, which further confirm USP14 effect of vimentin. In addition, we through bioinformatics analysis found that miR-320 a can target and down-regulate vimentin and USP14 at the same time, we further studied the effects of miR-320 a on gastric malignant behavior. Finally, we used the RT-qPCR detection the mRNA of USP14 and miR-320 a in gastric cancer tissues and normal gastric tissues and their correlation analysis, illustrating the miR-320 a can regulate and control vimentin and USP14 in gastric cancer cell. ResultRT-qPCR detection found that the vimentin expression were significantly increased when gastric cancer tissues compared to normal adjacent tissues, and MTT, plate clone formation experiment and Transwell experiments suggested that vimentin can accelerate the malignant behavior of gastric cancer cells. Ubiquitin experiment and Co-immunoprecipitation assays showed that USP14 interact with vimentin and USP14 can affect ubiquitination of vimentin. Morover, cell phenotype experiments proved that the USP14 also promotes malignant behavior of gastric cancer cells. In addition, we found that mi R-320 a contains the 3 ’UTR specific binding sites of vimentin and USP14, and found that miR-320 a impaired the expression of the two proteins. miR-320 a cell phenotype experiments aslo proved that the tumor suppressor role of miR-320 a in gastric cancer tissue. ConclusionIn this study, we found that vimentin was upregulated in human gastric cancer(GC) tissues and which can promote cell growth, migration and invasion in GC cells. Further study found that Ubiquitin-specific protease 14(USP14) can inhibit the ubiquitin level of vimentin to increase its expression in GC cells. In addition, we found that miR-320 a can regulate the expression of vimentin in transcription level. Research proves that miR-320 a can not only through target and downregulated vimentin directly, which also can bind and inhibit USP14 to reduce the expression of vimentin indirectly in gastric cancer cells. Moreover, miR-320 a can deregulate USP14 and vimentin to inhibit the malignant behavior of GC cells. Taken together, these results provide new insight into malignancy in gastric cancers. |
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