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Objective The study explored the effect of the knockout of the NCAPH gene on the proliferation, migration and invasion of pancreatic cancer cells, using NCAPH knockout pancreatic cancer cells generated by CRISPR/Cas9 technology. Methods The expression levels of NCAPH were analyzed in multiple cancer tissues and normal tissues by the Oncomine database.Kaplan-Meier analysis was used to investigate the correlation between the expression of NCAPH and the survival of pancreatic cancer patients. NCAPH-knockout cells using lentivirus were produced using the CRISPR/Cas9 technique. The proliferation-related molecules MEK and ERK were determined by Western blotting. CCK-8, colony formation, wound healing and Transwell assay were adopted to detect cell proliferation, migration and invasion of PANC cells. Western blotting detects the expression of MEK, p-MEK, ERK and p-ERK proteins. Results The results showed that NCAPH was significantly upregulated compared with paracancerous tissues in multiple cancer. Kaplan-Meier survival analysis revealed that patients with high expression of NCAPH were associated with a worse prognosis. The Western blotting showed that CRISPR/Cas9 technology efficiently disrupted the NCAPH gene and inhibited its expression in PANC cells. CCK-8 assay and colony formation assay showed that, compared with the control group, inhibiting the expression of NCAPH can inhibit the cell proliferation at 48, 72, 96 and 120 h (0.488±0.007 vs. 0.411±0.004,0.689±0.004 vs. 0.497±0.010, 1.071±0.034 vs. 0.689±0.020, 1.441±0.038 vs. 0.855±0.025) and the colony formation ability(210.0±2.9 vs. 144.0±16.4), the difference was statistically significant (P<0.05). Cell scratch and Transwell invasion assay showed that, compared with the control group, NCAPH knockout significantly suppressed cell migration (34.9%±1.7% vs. 15.1%±2.1%)and invasion [(351±23.64) cells vs. (194±13.0) cells], the difference was statistically significant (P<0.05). Then, we investigated the molecular mechanisms of this change, compared with the control group, NCAPH knockout significantly inhibited the expression of p-MEK and p-ERK proteins. Conclusion NCAPH knockout might inhibit proliferation, migration and invasion of PANC cells via the MAPK-ERK signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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