Construction of specific Smo lentivirus and expression of infected pancreatic cancer cells positive for CD24CD44 surface antibody.

This study aimed to construct a vector lentivirus carrying the Smo gene and transfect pancreatic cancer cells positive for CD24CD44 surface antibody and detect the infectivity. A lentivirus carrying a specific Smo fragment was designed and synthesized, and its functionality was tested. An overexpression group, inhibitory group, and negative control group were used for subsequent experimental research and comparison. A virus was successfully designed and produced. The best viral load was the 1X106 TU virus, where the cell growth and fluorescence effect of culture wells with polybrene dilution were the best. These are the transfection conditions and transfection param-eters for subsequent experiments. This plasmid was detected with a flag antibody by Western blot. The result was that it had a large specific 250kD band, and the membrane protein was overexpressed successfully. The expression results of Smo in five groups of cells after virus transfection detected by RT-PCR: blank group were 1.0038±0.0344, CON238 negative group: 1.0276±0.2944d, CON077 negative group: 0.8793±0.0402; LV-SMO15570-2 overexpres-sion group: 2.7479±0.8308, and LV-SMO-RNAi37304-1 inhibition group: 0.2386±0.0481. There were differences among the overexpression group and inhibition group with the other three groups. Homogeneity of variance: Bartlett F = 4.3530, P = 0.0016 < 0.05, heterogeneous. K-W test: cc2 = 10.9905* P = 0.0267, and there was a statisti-cally significant difference. The designed virus achieved the goal requirements. An sRNA fragment was designed for the key gene Smo of the Hh signaling pathway, and a vector lentivirus carrying this fragment was successfully constructed. The expression of Smo was analyzed after transfecting SW1990CD24CD44 positive cells, suggesting that the function of the RNA fragment designed for the key gene Smo in this experiment was successful.