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Objective To investigate the effects of silencing axon guide protein 4D (Sema4D) on the growth, autophagy and epithelial-mesenchymal transformation of gastric cancer cell SGC-7901. Methods The cells were randomly divided into control group, shRNA-NC group, Sema4D-shRNA1 group, Sema4D-shRNA2 group and Sema4D-shRNA3 group, and transfected into SGC-7901 with shRNA Sema4D vector. The expression of Sema4D mRNA was detected by RT-PCR. The clone formation rate was determined by colony formation assay. Flow cytometry was used to detect apoptosis. Morphological changes of epithelial-mesenchymal transformation were observed microscopically. Western blotting was used to detect the expression levels of Ki67, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, caspase-3, cleaved caspase-3, Beclin1, p62, LC3Ⅰ, LC3Ⅱ, E-cadherin, N-cadherin, and vimentin proteins. Results The results of RT-PCR showed that compared with the control group, the mRNA levels of Sema4D-shRNA1 group, Sema4D-shRNA2 group and Sema4D-shRNA3 group were significantly lower, and the mRNA level of Sema4D-shRNA3 group was significantly lower than that of Sema4D-shRNA1 group and Sema4D-shRNA2 group (P<0.05).Sema4D-shRNA3 was selected for subsequent experiments. The results of colony forming assay showed that the colony forming efficiency of Sema4D-shRNA3 group was significantly lower than that of the control group (P<0.05). Flow cytometry results showed that compared with the control group, the apoptosis rate of Sema4D-shRNA3 group was significantly increased (P<0.05).Western blotting results showed that compared with the control group, the expression levels of Ki67, PCNA, p62, N-cadherin and vimentin proteins in the Sema4D-shRNA3 group were significantly lower, Beclin1 and E-cadherin proteins were significantly higher, and the ratios of Bax/Bcl-2, cleaved caspase-3/caspase-3 and LC3Ⅱ/LC3Ⅰ were significantly higher (P<0.05). Microscopically, the epithelial-mesenchymal transformation was inhibited in the Sema4D-shRNA3 group. Conclusion shRNA Sema4D can inhibit proliferation, autophagy and epithelial-mesenchymal transformation of SGC-7901 cells and promote apoptosis.
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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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