| Purpose: This topic mainly studies the effect of Sinoporphyrin sodium mediated(DVDMS)photodynamic therapy(PDT)on long noncoding RNA-lnc-NPIPB11-1 and endoplasmic reticulum stress and lncNPIPB11-1 and its function in colorectal cancer cells,and investigated the role of lnc-NPIPB11-1 in DVDMS-PDT-induced endoplasmic reticulum stress in colorectal cancer cells.Methods:1.The microarray data of colorectal cancer CX-1 cells were processed by DVDMS-PDT,and the target gene lnc-NPIPB11-1 was screened out,and GO and KEGG functional enrichment analysis was performed.2.The cytotoxicity of DVDMS-PDT on colorectal cancer HT29 cells was detected by CCK-8 cytotoxicity assay.3.The expression of lnc-NPIPB11-1 in colorectal cancer HT29 cells treated with DVDMS-PDT and xenografted tumor were detected by q RTPCR,q RT-PCR and WB were used to detect the expression of endoplasmic reticulum stress molecules BIP and PERK in HT29 cells treated with DVDMS-PDT.4.Use the UCSC Human Genome Explorer to view the genome mapping of lnc-NPIPB11-1 and to predict the coding ability of lncNPIPB11-1 from the LNCipedia and CPC databases.The subcellular localization of lnc-npipb 11-1 was determined by lnc Locator database and nuclear/cytoplasmic segregation assay.5.HT29 and RKO cells were treated with low knock-down and overexpression of lnc-NPIPB11-1.The effects of lnc-NPIPB11-1 on BIP and PERK were examined by q RT-PCR and WB.6.The effects of lnc-NPIPB11-1 on the progression of colorectal cancer cells were examined by CCK-8 proliferation test,colony formation test and scratch test in vitro7.The effect of LNC-NPIPB11-1 on tumor progression was verified by subcutaneous xenotransplantation in vivo.8.BIP expression was detected in human colorectal cancer tissue and subcutaneous tumor tissue of nude mice by Shengxin data analysis9.The effects of lnc-NPIPB11-1 on colorectal cancer treated by DVDMS-PDT were tested by CCK-8 toxicity test and WBResults:1.The microarray data of colorectal cancer CX-1 cells treated with DVDMS-PDT showed that the expression of lnc-NPIPB11-1 was significantly higher than that of the untreated group,go and KEGG functional enrichment analysis indicated that lnc-NPIPB11-1 might be associated with ER stress.2.DVDMS-PDT inhibited the activity of HT29 cells in a time-and concentration-dependent manner3.In HT29 cells treated with DVDMS-PDT,the expression of lncNPIPB11-1,BIP and PERK also increased in a time-and concentrationdependent manner.4.LNC-NPIPB11-1,located on the chromosome 16 of the human genome(CHR16:29262273-29264479),is expressed in both the nucleus and cytoplasm,and is highly expressed in colorectal cancer.5.The expression of BIP and PERK decreased significantly when lncNPIPB11-1 was knocked down,and increased significantly when LNCNPIPB11-1 was overexpressed.6.The ability of cell proliferation and migration was decreased when lnc-NPIPB11-1 was knocked down,and the ability of cell proliferation and migration was enhanced when LNC-NPIPB11-1 was overexpressed.7.When lnc-NPIPB11-1 was knocked down,the size and weight of the tumors were lower than that of the control group,and when lncNPIPB11-1 was overexpressed,the size and weight of the tumors were higher than that of the control group.8.Bioinformatics data and q RT-PCR showed that the expression of BIP in colorectal cancer was higher than that in para-cancer tissue.Furthermore,the expression of BIP in the subcutaneous tumors of low-lncNPIPB11-1 knockout nude mice was significantly lower than that of the control group,while BIP was significantly higher in the subcutaneous tumors of overexpressing lnc-NPIPB11-1 than that of the control group.9.After silencing of lnc-NPIPB11-1,PDT treatment with DVDMSPDT decreased the cell viability,and the expression of BIP and PERK increased compared with the silencing group,but decreased compared with the DVDMS-PDT group,after treatment with DVDMS-PDT,the cell viability was restored,and the expression of BIP and PERK was increased compared with over-expression group,and also increased compared with DVDMS-PDT group,but there was no significant difference.Conclusion: lnc-NPIPB11-1 may act as a proto-oncogene targeting endoplasmic reticulum stress molecules BIP and PERK,thus promoting the occurrence and development of colorectal cancer,and endoplasmic reticulum stress may antagonize the killing effect of DVDMS-PDT on colorectal cancer. |
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