Application of human CDC37L1 gene and protein encoded by human CDC37L1 gene in preparation of medicine for treating gastric cancer
A gene encoding, protein technology, applied in the application field of medicine
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[0105] 3) Preparation of BSA standard curve.
[0106] 4) Take 100 μL of the diluted BSA standard solution and add it to a 96-well microplate, and take 2 parallel samples for each concentration.
[0107] 5) After adding 100 μL of working solution to each well, mix immediately.
[0108] 6) After reacting in a water bath at 37°C for 30 minutes, cool to room temperature.
[0109] 7) Use a spectrophotometer to measure the absorbance value at 562 nm. All samples should be tested within 20 minutes as much as possible.
[0110] 8) The absorbance value of the BSA standard solution of each concentration is subtracted from the average value of the Blank value, and the standard curve of the BSA standard solution is drawn.
[0111] 9) Take 100 μL of the dilution solution of the sample to be tested and add it to a 96-well microwell plate, and take 2 parallel samples for each sample.
[0112] 10) After adding 100 μL of working solution to each well, mix immediately.
[0113] 11) After r...
Embodiment 1
[0128] Silencing CDC37L1 expression promotes the proliferation of gastric cancer cells
[0129] Through NCBI online search (https: / / www.ncbi.nlm.nih.gov / ), the cDNA sequence CCDS6454.1 of CDC37L1 was retrieved ( figure 1 A) in and its coded amino acid sequence NP_060383.2 ( figure 1 in B).
[0130] First, gastric cancer cells were transfected with exogenous CDC37L1+3Flag plasmid and its control plasmid, and CDC37L1 was successfully overexpressed through Western Blot experiments; then gastric cancer cells were transfected with siCDC37L1 targeting CDC37L1 and control siRNA, and small interfering RNA was tested by Western Blot The effect on the expression of CDC37L1 in gastric cancer cells, the results showed that the expression of CDC37L1 protein was significantly decreased ( figure 2 in A).
[0131] Gastric cancer MGC-803 cells in the logarithmic growth phase were seeded in 24-well plates, and the effect of CDC37L1 on the proliferation of gastric cancer cells was tested by...
Embodiment 2
[0133] CDC37L1 inhibits the growth ability of gastric cancer cells
[0134] The functions of long-term and short-term CDC37L1 on the growth of gastric cancer cells were detected by plate colony formation and CCK8 growth curve experiments, respectively.
[0135] Gastric cancer cells (control group LV-NC and LV-shNC) were infected with overexpression lentivirus LV-CDC37L1 and knockdown lentivirus LV-shCDC37L1 respectively, and the cell lines with stable expression were obtained by Puromycin screening, and the cells in the logarithmic growth phase were collected in each well. 2*10 3 Cells were seeded in a six-well plate, and plate colony formation experiments were performed. After 2-3 weeks of culture, the effect of CDC37L1 on the long-term growth of gastric cancer cells was detected. The statistical results showed that down-regulation of CDC37L1 significantly promoted the growth of gastric cancer cells ( image 3 in A).
[0136] Gastric cancer cells were transiently transfecte...
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