Molecular marker MLLT11 for diagnosing and treating bladder cancer and application of molecular marker MLLT11
A technology of bladder cancer and markers, which is applied in the target and application fields of bladder cancer proliferation and metastasis research, and can solve the problems of lack of predictive indicators for bladder cancer and failure to improve the survival rate of bladder cancer patients
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Embodiment 1
[0052] Example 1, MLLT11 is up-regulated in bladder cancer patients and bladder cancer cell lines
[0053] 1. Tissue samples
[0054] Tissue samples from 33 patients undergoing bladder cancer surgery were collected from March 2014 to December 2015, provided by the Urology Department of the First Affiliated Hospital of Wenzhou Medical University. Collect patient name, gender, age, pathology number, pathology and other information. Most of the selected patients were male and all underwent total cystectomy. Preoperative biopsy and postoperative pathological examination confirmed bladder urothelial carcinoma. Tumor tissue was collected from each sample, and normal tissue was cut about 3 cm away from the tumor as a control. Each sample was frozen in liquid nitrogen after the tissue was isolated, and the experimental operation was approved by ethical review.
[0055] 2. Immunohistochemistry
[0056] Refer to the immunohistochemical method, as follows:
[0057] 2.1. Baking slice...
Embodiment 2
[0088] Example 2. Analysis of TCGA database shows that the upregulation of MLLT11 expression is negatively correlated with the 5-year DFS and OS of bladder cancer patients
[0089] In order to clarify the possible role of MLLT11 in bladder cancer and the relationship with bladder cancer patients, the present invention analyzed the 5-year disease-free survival (DFS) and overall survival (OS) of bladder cancer patients in the TCGA database and the relationship between MLLT11 and MLLT11. The relationship between the amount of expression ( figure 2 A-2B), the median expression level of MLLT11 in bladder cancer tissues was used to divide them into two groups with high and low expression. The results showed that patients with low MLLT11 expression had longer 5-year DFS and 5-year OS and higher survival rate; on the contrary , patients with high expression of MLLT11 had shorter 5-year DFS and 5-year OS and lower survival rate, and there was a significant difference between the two g...
Embodiment 3
[0091] Example 3. Knocking out the expression of MLLT11 can significantly inhibit the proliferation ability of bladder cancer cells in vitro
[0092] 1) First, two commonly used bladder cancer cell lines T24T and UMUC3 were selected to construct MLLT11 knockout stable cell lines, and the Cas9 vector, MLLT11 KO#1, MLLT11 KO#2, and MLLT11 KO#3 plasmids were transfected into T24T and UMUC3 respectively Cells, stable cell lines obtained by monoclonal screening, western blot detection of MLLT11 protein level to confirm the knockout efficiency. The results showed that both T24T and UMUC3 cells successfully knocked out the expression of MLLT11 protein, and GAPDH was used as an internal reference ( Figure 3-1 ).
[0093] The sequence of MLLT11 KO#1 is shown in SEQ ID NO.1; the sequence of MLLT11 KO#2 is shown in SEQ ID NO:2; the sequence of MLLT11 KO#3 is shown in SEQ ID NO:3.
[0094] 2), soft agar colony formation experiment
[0095] A soft agar colony formation experiment was c...
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