Lnczc3h7a inhibits proliferation and migration of intestinal cancer cells by targeting CTHRC6
Objective:To investigate the effect of Lnczc3h7a on colorectal cancer cell proliferation and migration to provide a potential target for colorectal cancer treatment.Methods:Colorectal cancer cell lines (HT-29, SW-60, HCT-116, Lovo, COLO320DM, and DLD-1) and normal colorectal epithelial cell line (FHC) were used in this study, and the expression of Lnczc3h7a in these cell lines was detected by fluorescence quantitative PCR. HCT-116 cells were divided into three groups and transfected with Lnczc3h7a mimic, Lnczc3h7a inhibitor, and empty plasmid, respectively. The proliferation and migration of the three groups of cells were detected and compared by the CCK8 method and scratch assay, respectively. The effect of Lnczc3h7a on CTHRC6 protein expression was detected by Western blot after knockdown of Lnczc3h7a using the CRISP-Cas9 technique.Results:The expression levels of Lnczc3h7a in HT-29, SW-60, HCT-116, Lovo, COLO320DM, and DLD-1 cells were 0.64±0.02, 0.67±0.03, 0.59±0.04, 0.68±0.03z, 0.72±0.04, and 0.62±0.05, respectively, which were significantly lower than that in normal colorectal epithelial cells (FHC; 1.00±0). After transfection, the expression level of Lnczc3h7a in the Lnczc3h7a mimic group was (1.36±0.05), significantly higher than that in the Lnczc3h7a inhibitor group (0.57±0.04) and the blank control group (1.02±0.03) (P＜0.05). After transfection for 1~3 days, the difference in cell proliferation was not significant among the three groups; on the 4th and 5th days of transfection, the proliferation of colorectal cancer cells in the Lnczc3h7a inhibitor group (0.84±0.06 and 1.08±0.06) was significantly higher than that in the blank control group (0.73±0.06 and 0.96±0.06) and the Lnczc3h7a mimic group (0.67±0.05 and 0.91±0.04), and the proliferation of colorectal cancer cells in the blank control group was higher than that in the Lnczc3h7a mimic group (P＜0.05). At 24 h and 48 h of cell culture, the migration of colorectal cancer cells in the Lnczc3h7a inhibitor group (447.96±46.19 and 467.23±45.36) was higher than that in the control group (416.57±59.48 and 425.17±58.34) and the Lnczc3h7a mimics group (416.57±59.48 and 425.17±58.34), and the migration ability of colorectal cancer cells in the control group was higher than that in the Lnczc3h7a mimic group (all P＜0.05). After knockdown of Lnczc3h7a using the CRISP-Cas9 technique, RT-PCR showed that the expression of Lnczc3h7a was substantially decreased at the mRNA level (P＜0.01), while Western blot analysis showed that there was no expression of Lnczc3h7a protein. Western blot analysis showed that the Lnczc3h7a inhibitor group had significantly higher CTHRC6 protein expression levels than the blank control group (P＜0.05), and CTHRC6 protein expression levels were significantly decreased in the Lnczc3h7a mimic group compared to the control group (P＜0.05), while the Lnczc3h7a knockout group had significantly higher CTHRC6 expression than other groups (P＜0.05).Conclusion:The expression of Lnczc3h7a is reduced in colorectal cancer cell lines. Overexpression of Lnczc3h7a could inhibit the proliferation and migration of colorectal cancer cells. Lnczc3h7a inhibits the growth and metastasis of colorectal cancer possibly by targeting CTHRC6 expression.
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