Tumor therapy medicine targeting SCNN1A and application
A drug and tumor technology, applied in anti-tumor drugs, gene therapy, drug combination, etc., can solve problems such as unsatisfactory treatment effect and prognosis, and endanger human health
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Embodiment 1
[0027] Example 1 Expression Analysis of SCNN1A in Tumor Tissue
[0028] PCR technology was used to detect the expression of SCNN1A in human colon cancer and its adjacent cancer, liver cancer and its adjacent cancer, lung cancer and its adjacent cancer. The cases in each group included 355 cases of colon cancer tissue, 19 cases of colon cancer tissue and 13 cases of liver cancer tissue. , 7 cases of liver cancer tissue; 112 cases of lung cancer tissue, 10 cases of lung cancer tissue. The results showed that the expression of SCNN1A in colon cancer, liver cancer, and lung cancer tissues was significantly lower than that in adjacent tissues, such as figure 1 a.
[0029] Further verification in human liver cancer cells: extract the mRNA of human normal liver cell line LO2, liver cancer cell lines Hep3B, Bel7407, Huh7, HepG2, and use real-time fluorescent quantitative PCR to detect the expression level of SCNN1A, the results show that Hep3B, Bel7407, Huh7 and other cell lines The...
Embodiment 2
[0033] Example 2 In vitro inhibitory effect of SCNN1A on liver cancer cells Hepa1-6
[0034] The cDNA sequence of the murine SCNN1A gene was constructed into the eukaryotic overexpression backbone vector pcDNA3.1, and the Hepa1-6 cell line was transfected by liposome method, and the pcDNA3.1 empty vector was transfected at the same time as a control. G418 screened to obtain resistant clones, and then detected the expression of SCNN1A by real-time quantitative PCR, and selected SCNN1A-pcDNA3.1#1 clones with similar expression levels of SCNN1A in liver tissue and pcDNA3.1#1 with similar expression levels to Hepa1-6 for in vitro experiments . Among them, quantitative results such as figure 2 a.
[0035] MTT, cell scratch, colony and flow cytometry were performed on Hepa1-6, pcDNA3.1-Hepa#1 (referred to as pcDNA3.1), SCNN1A-pcDNA3.1-Hepa#1 (referred to as SCNN1A-pcDNA3.1) cell lines respectively The cycle assay detects the growth inhibition of SCNN1A on the Hepa1-6 cell line. ...
Embodiment 3
[0040] Example 3 Subcutaneous tumor-bearing results of nude mice overexpressing Hepa1-6 in SCNN1A
[0041] 15 male nude mice were randomly divided into 3 groups, 5 in each group, and each mouse was inoculated with 6×10 6 The three groups of cell lines SCNN1A-pcDNA3.1, pcDNA3.1, and Hepa1-6 were inoculated subcutaneously in the left upper limb of nude mice respectively. Tumor formation was observed every day after inoculation. From the 7th day after injection, the tumor volume was measured according to the prescribed time. A tumor with a diameter of 3 mm was regarded as a tumor, and the tumor size was recorded. The longest and shortest diameters of the transplanted tumors were measured with a vernier caliper, once every 3 days, and the tumor volume was calculated according to the formula "tumor volume V = long diameter × short diameter × short diameter / 2". According to the average tumor volume, the tumor growth curve was drawn. On the 15th day, the nude mice were sacrificed...
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