Expression and regulatory factor analysis of tumor associated calcium signaling sensor 2 in colorec-tal cancer
Objective To investigate the expression changes of tumor associated calcium signaling sensor 2 in colorectal cancer tissues and the regulation of its expression levels by upstream microRNA(miRNA,miR).Methods Totally,58 cases of clinical specimens and adjacent tissues of colorectal cancer admitted to the First Affiliated Hospital of Xinxiang Medical College from January 2020 to January 2023 were selected as the research objects.The differentially expressed proteins between colorectal cancer and adjacent tissues were analyzed by two-dimensional electrophoresis.Tumor related calcium signal sensor 2 was selected as the research object,and the expression levels of tumor related calcium signal sensor 2 in adjacent tissues and colorectal cancer tissues were analyzed by Western blotting.The positive rates of tissue cell proliferation antigen proliferating cell nuclear antigen(PCNA)in two groups were analyzed by immuno-histochemistry.The relationship between tumor related calcium signal sensor 2 and cell proliferation was analyzed.miRNA transcriptomics in adjacent and colon cancer tissues was analyzed by transcriptomic anal-ysis and screened for differentially targeted miRNAs for tumor associated calcium signaling sensor 2.The targeting of miRNAs was analyzed by dual luciferase reporter gene.The comparison of inter group measure-ment data was done by t-test.Results A total of 10 differentially expressed protein components were screened by two-dimensional electrophoresis.In this study,the protein tumor related calcium signal sensor 2 with significant differences was selected as the research object.The protein expression level of tumor re-lated calcium signal sensor 2 in adjacent tissues(0.92±0.14)was significantly lower than that in colorec-tal cancer tissues(1.62±0.20)(t=14.320,P＜0.05).The protein expression level of tumor related calcium signal sensor 2 in patients with negative PCNA expression(1.33±0.13)was significantly lower than that in colorectal cancer tissue(1.69±0.16)(t=6.092,P＜0.05).There was a total of 1 218 dif-ferentially expressed miRNAs between adjacent and colorectal cancer tissues,of which 732 were upregulated and 486 were downregulated.There were 5 complementary miRNAs which were found to be associated with tumor associated calcium signaling sensor 2,namely miR-488-3p,miR-625-5p,miR-3650,miR-518a-5p,and miR-204-3p.The expression levels of miR-488-3p,miR-625-5p,miR-3650,miR-518a-5p,and miR-204-3p in adjacent tissues(1.07±0.08,1.07±0.009,1.11±0.15,1.01±0.10,1.15±0.15)were significantly higher than those in colorectal cancer tissues(0.28±0.05,0.30±0.07,0.45±0.08,0.54±0.09,0.69±0.12)(t=21.050,26.350,14.850,12.600,9.165,P＜0.05).The luciferase activity in cells co transfected with miRNA control and luciferase reporter genes(0.96±0.07,0.94± 0.03,0.92±0.06,0.91±0.08,0.95±0.05)was significantly higher than that in cells co transfected with miR-488-3p,miR-625-5p,miR-3650,miR-518a-5p,miR-204-3p mimetics and luciferase reporter genes(0.22±0.06,0.24±0.07,0.34±0.09,0.45±0.08,0.60±0.12)(t=23.541,23.102,18.208,14.231,12.514,P＜0.05).Conclusion Tumor associated calcium signaling sensor 2 is significantly up-regulated in colorectal tissue and is regulated by miRNAs such as miR-488-3p,miR-625-5p,miR-3650,miR-518a-5p,and miR-204-3p.
Tumor related calcium signal sensor 2Colorectal cancerProteomicsMicroRNAsTranscriptomics
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