Application of wdr1 gene in preparation of non-small cell lung cancer treatment and detection products
A non-small cell lung cancer and gene technology, applied in the field of molecular biology, can solve the problems that have not been reported in the research, achieve the effect of improving the current treatment status and good application prospects
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Embodiment 1
[0028] Example 1 WDR1 expression difference in NSCLC cancer tissue and its normal tissue
[0029] In this example, the total RNA of NSCLC cancer tissue and its normal tissue was extracted, and the WDR1 fragment gene and the internal reference gene b-actin were respectively amplified by real-time fluorescent quantitative PCR, and the expression difference of WDR1 gene in NSCLC cancer tissue and normal tissue was analyzed. The specific method includes the following steps:
[0030] (1) Collection of NSCLC cancer tissue and its normal tissue samples: In this example, NSCLC cancer tissue and its normal tissue were used as the detection objects. The samples of 39 NSCLC patients were collected from Nanjing Chest Hospital of Southeast University; Approved by the Medical Ethics Committee of Nanjing Chest Hospital, all cases obtained the informed consent of the patients;
[0031] (2) Extract total RNA from the NSCLC cancer tissue and its normal tissue to be tested: add Trizol (Invitrog...
Embodiment 2
[0037] Example 2 Evaluating the relationship between the expression level of WDR1 gene and the prognosis of NSCLC patients
[0038] In order to evaluate the relationship between the expression level of WDR1 and the prognosis of patients with non-small cell lung cancer, we performed Kaplan-Meier survival analysis through an online tool (http: / / kmplot.com) on a cohort of about 960 patients with non-small cell lung cancer Analysis shows (such as figure 2 shown), high WDR1 gene expression levels were associated with decreased overall survival (HR=1.43, log-rankP=3.7E-08). In addition, we also used another systematic software to evaluate the relationship between WDR1 gene expression level and patient prognosis (http: / / www.oncolnc.org), and we also obtained similar results (results such as image 3 shown): High levels of WDR1 gene expression were associated with reduced survival in lung adenocarcinoma (P=0.0428) and lung squamous cell carcinoma (P=0.193). The above results indica...
Embodiment 3
[0039] Example 3 Knocking down WDR1 at the cellular level to study the role of WDR1 in non-small cell lung cancer
[0040] We first adopted the method reported by Moffat J., et al., 2006, using the pLKO lentiviral expression system to construct stable knockdown WDR1 cells (shWDR1) and control cells in two non-small cell lung cancer cell lines A549 and H1299 (shCTL); the shRNA sequence we designed and used was as follows: shWDR1-GCTGGGAAGATCAAAGACATT (as shown in SEQ ID NO: 6); shCTL: GCAAGCTGACCCTGAAGTTCAT. The A549 cell lines in the WDR1 stable knockdown group and the control group were screened by lentivirus packaging, infection and antibiotic puromycin, and finally the cell selection was verified by Western blot. The Western blot experiment method is as follows: Cell protein was extracted by RIPA cell lysate, protein concentration was determined by BCA method, the same amount of protein samples were subjected to 10% SDS-PAGE electrophoresis, transferred to the membrane, blo...
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