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| The effects of knockdown AGK gene on proliferation and migration capacity of colorectal cancer cells |
| Objective In order to investigate the effects of acylglycerol kinase(AGK) on proliferation and migration in HCT116
cells. Methods In this study, shRNA silencing lentivirus plasmid of AGK gene was constructed, and the second generation lentivirus
packaging system was used to prepare and concentrate lentivirus venom and infect human colon cancer cell HCT?116 cell line.Then
HCT?116 cell lines with knockdown AGK gene were screened for puromycin resistance.qRT?PCR and Western blotting were used
to detect the mRNA and protein expression of AGK after knockdown. CCK?8 kit was used to detect the proliferation of AGK
knockdown stable cell lines.The effect of AGK knockout on cell migration was detected by cells scratch assay. Results ShRNA
silencing lentivirus plasmid of AGK gene constructed in this experiment can effectively knockdown endogenous AGK in HCT116.The
mRNA and protein levels of AGK in HCT116?ASR?876 and HCT116?ASR?878 cells were significantly lower than those of wild?type
cell lines (P<0.001).CCK?8 detection showed that AGK knockdown significantly reduced cell proliferation ability (P<0.05).The scratch
test showed that AGK knockdown could significantly inhibit cell migration. Conclusion Knockout of AGK gene can significantly
reduce the proliferation and migration abilities of HCT116 cells, which might provide a new potential target for targeted therapy of
colorectal cancer. |
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Abstract Objective In order to investigate the effects of acylglycerol kinase(AGK) on proliferation and migration in HCT116
cells. Methods In this study, shRNA silencing lentivirus plasmid of AGK gene was constructed, and the second generation lentivirus
packaging system was used to prepare and concentrate lentivirus venom and infect human colon cancer cell HCT⁃116 cell line.Then
HCT⁃116 cell lines with knockdown AGK gene were screened for puromycin resistance.qRT⁃PCR and Western blotting were used
to detect the mRNA and protein expression of AGK after knockdown. CCK⁃8 kit was used to detect the proliferation of AGK
knockdown stable cell lines.The effect of AGK knockout on cell migration was detected by cells scratch assay. Results ShRNA
silencing lentivirus plasmid of AGK gene constructed in this experiment can effectively knockdown endogenous AGK in HCT116.The
mRNA and protein levels of AGK in HCT116⁃ASR⁃876 and HCT116⁃ASR⁃878 cells were significantly lower than those of wild⁃type
cell lines (P<0.001).CCK⁃8 detection showed that AGK knockdown significantly reduced cell proliferation ability (P<0.05).The scratch
test showed that AGK knockdown could significantly inhibit cell migration. Conclusion Knockout of AGK gene can significantly
reduce the proliferation and migration abilities of HCT116 cells, which might provide a new potential target for targeted therapy of
colorectal cancer.
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Received: 10 October 2020
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Corresponding Authors:
shihanping
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