Gastric adenocarcinoma marker Drosha protein and applications thereof
A gastric adenocarcinoma and protein technology, applied in the field of gastric adenocarcinoma marker Drosha protein, achieves the effect of important clinical application value, simple application method, high specificity and sensitivity
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Embodiment 1
[0040] Example 1 Correlation analysis between the expression of Drosha protein in cytoplasm and nucleus and the clinicopathology of gastric adenocarcinoma
[0041] Materials: 829 tissue microarray samples of human gastric adenocarcinoma in different pathological stages.
[0042] Methods: The tissue chips of 829 cases of gastric adenocarcinoma were analyzed by immunohistochemical staining, and IPP software was used to detect the expression of Drosha in the cytoplasm and nucleus of the tissue chips.
[0043] Immunohistochemical staining
[0044] 1) Dewaxing: For paraffin-embedded tissue chips, first melt the paraffin in a 60°C incubator, and then put them in xylene I, II, III for 10 minutes respectively;
[0045] 2) Hydration: put the dewaxed tissue chip into 100% alcohol I, II, 95% alcohol, 85% alcohol, 70% alcohol, PBS buffer I, II, III in sequence, each for 5 minutes;
[0046] 3) Block endogenous peroxidase: add 3% H2O2 dropwise to cover the tissue, act at 37°C for 10 minut...
Embodiment 2
[0064] Example 2 Inhibition of the expression of Drosha promotes the invasion of gastric cancer cells
[0065] Material: Gastric cancer cell line SGC-7901.
[0066] Methods: Western blot was used to detect the expression of Drosha protein in SGC-7901 cells before and after siRNA interference.
[0067] Cell culture and cell transfection
[0068] SGC-7901 cells were routinely cultured in RPMI1640 medium containing 10% fetal bovine serum. When the cell fusion was about 60%, according to Roche liposome transfection instructions, siRNA and negative control sequence were transfected into SGC-7901 cells respectively, and protein was collected 48 hours after transfection for detection and Transell invasion experiment.
[0069] In order to further verify the correlation between Drosha and the depth of tumor invasion in Table 1 of Example 1, the expression of Drosha in the gastric cancer cell line SGC-7901 was suppressed with siRNA (see figure 2 ), and then Transwell detected the in...
Embodiment 3 25
[0070] Example 3 Correlation analysis of the prognosis of 225 patients with gastric adenocarcinoma
[0071] In order to further study the relationship between the cytoplasmic expression of Drosha and the prognosis of patients with gastric adenocarcinoma, the relevant prognosis analysis was performed on the 225 patients who were predicted in Example 1. Such as Figure 4 As shown, the Kaplan survival curve analysis showed that the expression level of cytoplasmic Drosha was significantly correlated with the prognosis of adenocarcinoma patients (LR=7.088, P=0.008), while the expression level of nuclear Drosha was not correlated with the prognosis of adenocarcinoma patients (LR=1.954, P=0.162).
[0072] In order to further explore whether cytoplasmic / nuclear Drosha can be used as an independent prognostic factor for patients with gastric adenocarcinoma, COX univariate analysis was used to find out the potential prognostic factors related to the prognosis of patients. As shown in ...
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