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MiR-411 serving as target of bladder cancer and application of miR-411

A technology of mir-411, 1.mir-411, applied in miR-411 as the target of bladder cancer and its application field, can solve the problems that bladder cancer-related functions have not been reported, and achieve the effect of enhancing sensitivity

Inactive Publication Date: 2016-10-12
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above studies show that the biological function of miR-411 is significantly organ-tissue dependent, and its related function in bladder cancer has not been reported

Method used

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  • MiR-411 serving as target of bladder cancer and application of miR-411
  • MiR-411 serving as target of bladder cancer and application of miR-411
  • MiR-411 serving as target of bladder cancer and application of miR-411

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Experimental program
Comparison scheme
Effect test

experiment Embodiment 1

[0033] Establishment of animal model of bladder cancer induced by BBN

[0034] 1) Animal feeding

[0035] Purchase C57BL / 6 mice, 3-4 weeks old, and after 1 week of adaptive feeding, they are randomly divided into cages, with 4 mice in each cage. All animal experiments were conducted in accordance with the relevant regulations of the Ethics Committee of Wenzhou Medical University.

[0036] 2) Design of BBN treatment of mice in different weeks

[0037] Put the BBN (N-butyl-N-(4-hydroxybutyl) nitrosamine) substance into the drinking water of the mice in the animal room to make the final concentration 0.05%. After that, the drugs were given in turn, and the drinking water was changed twice a week, and all the mice were killed after the last week of administration. The general bladder tissue was taken from each mouse and embedded for immunohistochemistry, and the epithelial cells were scraped from the remaining half, and RNA was extracted after mixing every 4 mice in the same gr...

experiment Embodiment 2

[0049] miRNA expression profile microarray experiment (this part was completed by Hangzhou Lianchuan Biotechnology Co., Ltd.)

[0050] Microarray experiments were performed by LC Sciences. Microarray experiments used 4 μg of total RNA samples. A poly(A) tail is added to the 3' end of the total RNA by Poly(A) polymerase, and then ligated with an oligonucleotide label for subsequent fluorescent labeling. The hybridization reaction was carried out overnight on the μParaflo microfluidic chip using the circulation action of a microcirculation pump (Actic Technologies). On the chip, each detection probe is composed of a chemically modified nucleotide coding segment (complementary to the target microRNA (derived from miRBase, http: / / www.mirbase.org / ) or complementary to other RNAs (quality control or custom sequences) and a spacer composed of polyethylene glycol (enlarging the distance between the coding segment and the substrate). The detection probes were synthesized in situ u...

experiment Embodiment 3

[0053] Expression of miR-411 in clinical bladder cancer samples and human bladder cancer cells

[0054] tissue sample

[0055] Tissue samples from 14 patients who underwent surgery for shoulder and wrist cancer in the Urology Department of the Second Affiliated Hospital of Wenzhou Medical University were collected. 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 3cm away from the tumor as a control. Each sample was frozen in liquid nitrogen after tissue isolation. The experimental operation was approved by ethical review.

[0056] Tissue / Cell Total RNA Extraction

[0057] a. Take out the bladder cancer clinical samples from the -80°C refrigerator, put about 30-50 μg of each sample in...

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Abstract

The invention belongs to the field of biotechnology, and specifically relates to miR-411 as a bladder cancer target and its application. The invention uses bladder cancer T24 and UMUC3 cells as models, and overexpressing miR-411 can inhibit the proliferation of bladder cancer cells in vivo and in vitro, and induce bladder cancer Cancer cell cycle G2 / M phase arrest, while enhancing the sensitivity of chemotherapy drugs (mitomycin and gemcitabine) to bladder cancer cells. It can be seen that miR-411 expression promoter can prevent or / and treat bladder cancer. Using miRNA chip technology, real-time fluorescent quantitative PCR technology, bioinformatics analysis of TCGA database and other methods to detect the expression of miR‑411 in bladder cancer primary animal models and clinical bladder cancer tissues showed a significant down-regulation trend, and the expression of miR‑411 was correlated with bladder cancer There is a certain positive correlation with the survival time of patients. It can be seen that by detecting the expression of miR‑411, bladder cancer can be assisted in the diagnosis and / or prognosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a new target of bladder cancer and its application, and more specifically relates to miR-411 as a bladder cancer target and its application. Background technique [0002] Bladder cancer is one of the most common malignant tumors, ranking first among urinary system malignant tumors. In recent years, environmental factors including smoking, exposure to chemical substances and bad living habits are closely related to the occurrence of bladder cancer and lead to a gradual increase in the incidence and mortality of bladder cancer. According to the latest published statistics, there are more than 350,000 new cases and 132,000 deaths worldwide each year. Despite its prevalence and socioeconomic impact, bladder cancer is the least studied tumor. The research and development of new targeted drugs and the application of new preventive measures are recognized as the main ways to cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7068C12Q1/68A61P35/00A61K31/407
CPCA61K45/00A61K31/407A61K31/7068C12Q1/6886C12Q2600/118C12Q2600/158
Inventor 黄海山黄传书高基民古佳艳许晓玲邓文海
Owner WENZHOU MEDICAL UNIV
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