Gastric cancer thermo-chemotherapy sensitive marker and application thereof
A gastric cancer and drug technology, applied in the field of molecular biology, can solve problems such as limited research, and achieve the effect of inhibiting proliferation, promoting apoptosis, and high accuracy
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Embodiment 1
[0061] Example 1 Screening of ZNF429 Gene Missense Mutations
[0062] Tumor tissue samples from 18 patients with gastric cancer with peritoneal metastasis were collected. With reference to HIPEC clinical research data at home and abroad, the survival time of gastric cancer patients after HIPEC treatment was greater than or equal to 11 months as good HIPEC efficacy, and the survival time less than 11 months was poor HIPEC efficacy , 18 patients with gastric cancer were divided into two groups with good curative effect and poor curative effect, and the baseline comparison between the two groups was carried out.
[0063] After statistical analysis, the results showed that the gender (P=0.999), ECOG score (P=0.999), pathological differentiation degree (P=0.999), peritoneal metastasis P scale (P=0.999), ascites volume (P=0.999) and ascites volume (P=0. =0.552) and the number of chemotherapy within half a year (P=0.751), there was no statistical difference (Table 1); then, the whole...
Embodiment 2
[0069] Example 2 ZNF429 Verification
[0070] This example shows that the expression of ZNF429 is up-regulated in gastric cancer tissues, and is associated with clinicopathological factors in gastric cancer patients.
[0071] 1. Using immunohistochemical method, the expression of ZNF429 in the cancer tissue and paracancerous tissue samples of 90 gastric cancer patients was detected, and the correlation analysis between the clinicopathological parameters and the expression of ZNF429 in 90 gastric cancer patients was further carried out. The specific operations were as follows:
[0072] Bake the tissue chip in a constant temperature oven at 60 degrees Celsius for 3 hours (see if the wax sheet has dissolved), followed by dewaxing and adsorption: 10 minutes in environmental protection clear agent 1, 10 minutes in environmental protection clear agent 2, hydration: absolute ethanol for 5 minutes , 95% ethanol for 5 minutes, 70% ethanol for 5 minutes, washed with PBS for 3 times, eac...
Embodiment 3
[0081] Example 3 ZNF429 Knockdown Verification
[0082] 1. Knockdown of ZNF429 can inhibit the viability of gastric cancer cells
[0083] By silencing endogenous ZNF429, qPCR detection found that siRNA of ZNF429 could significantly reduce the expression level of ZNF429 in gastric cancer cell lines MKN28, MKN45, and SNU1 (the cells were all purchased from Wuhan Punuosai Life Technology Co., Ltd.).
[0084] The specific operation of silencing endogenous ZNF429 is as follows:
[0085] The day before transfection, inoculate gastric cancer cells into a 6-well plate with a complete medium without antibiotics, and the inoculation density can ensure that the cell density reaches 50-80% at the time of transfection; Medium, wash with PBS or normal saline twice, then add 1.7mL serum-free Opti-MEM medium to each well; add transfection (siRNA2.5μg) in 150μL serum-free Opti-MEM medium, and mix gently , stand at room temperature for 5 minutes; gently mix Lipofectamine3000 before use, add 5...
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