| Objective:Colorectal cancer(CRC)is the fourth most deadly cancer with about 900000 deaths annually in the world.The main reason for its poor prognosis is distant metastasis,thus inhibiting or even blocking the metastasis is of supreme significance to prolong the survival time and improve the life quality of patients.Lnc RNAs(long non-coding RNAs)are involved in every stage of tumor metastasis and mitochondrial dynamics also play a vital role in this process.While few researchers focus on the relationship between Lnc RNA and mitochondrial dynamics in cancer.This study indicates that Lnc RNA SNHG11 may promote tumor metastasis by mediating mitochondrial dynamics.Exploring the potential molecular mechanism is of supreme significance to the advance of mitochondrial therapy and to reduce or even block the metastasis of colon cancer.Methods:1.Real-time fluorescence quantitative PCR(RT-PCR)was used to detect the expression of SNHG11 in Ls174 T,Caco2 and HCT-116 colorectal cancer cell lines.2.The overexpression plasmid SNHG11 and interfering plasmid sh-SNHG11,were designed and constructed,and the efficiency was verified by RT-PCR.3.The overexpression plasmid SNHG11 was transfected into Ls174 T,with relatively low expression of SNHG11,and the interfering plasmid sh-SNHG11 was transferred into HCT-116 cells,with relatively high expression of SNHG11.The changes of invasion and migration ability of colorectal cancer cells were detected by Transwell migration and invasion assays.The changes of morphology and location of mitochondria in colorectal cancer cells were observed by mitochondrial staining,and the changes of membrane potential were detected by membrane potential staining reagent,and the expression levels of mitochondrial fission and fusion proteins were detected by western blot.4.Western blot was used to investigate the effect of SNHG11 on FAK signaling pathway in colorectal cancer cells.Results:1.RT-PCR results showed that there was no statistical significance in the expression of SNHG11 among three kinds of colorectal cancer cell lines.The expression level of SNHG11 in Ls174 T was relatively low,while the expression level of SNHG11 in HCT-116 was relatively high.2.The results of Transwell invasion and migration assays showed that the upregulation of SNHG11 promotes the invasion and migration of colorectal cancer cells,while the down-regulation of SNHG11 inhibits the invasion and migration of colorectal cancer cells.3.The results of mitochondrial staining showed that the up-regulation of SNHG11 caused the mitochondria to become small and short fragment-like structures and move to the leading edge of the cells.The down-regulation of SNHG11 had no effect on the morphology of the mitochondria,while it made the mitochondria become more concentrated around the nucleus.4.The results of membrane potential detection showed that up-regulation of SNHG11 increased the membrane potential of cells,while down-regulation of SNHG11 decreased the membrane potential.5.The results of Western blot showed that up-regulation of SNHG11 promoted the expression of mitochondrial fission proteins such as Drp1,Fis1,MFF and p-Drp1(ser616),with no change of p-Drp1(ser637).While the expression of mitochondrial fusion proteins such as MFN1 and MFN2 was decreased,with no change of OPA1.After down-regulation of SNHG11,the expression of mitochondrial fission proteins such as Drp1,Fis1 and MFF was decreased,with no change of p-Drp1(ser616)and pDrp1(ser637)increased.While mitochondrial fusion proteins,for example,MFN1 decreased,with no change of MFN2 and OPA1.6.After up-regulation of SNHG11,the expression of FAK and p-FAK increased,while the expression of FAK and p-FAK was decreased after down-regulation of SNHG11.Conclusion:1.SNHG11 can promote the invasion and migration of colorectal cancer cells.2.SNHG11 can promote mitochondrial fission and inhibit mitochondrial fusion.3.SNHG11 can increase the cell mitochondrial membrane potential.4.SNHG11 may regulate the metastasis of colorectal cancer through FAK signaling pathway. |
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