Application of exosomal circular RNA 0109315 as a target in the prevention and treatment of benign prostatic hyperplasia

A technology for benign prostatic hyperplasia and exosomes, which is applied in the detection/testing of microorganisms, medical preparations containing active ingredients, and urinary system diseases, etc. question

Active Publication Date: 2022-03-08
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Regrettably, the low-throughput detection of 9 circRNAs selected for co-expression analysis (circ_0006869, circ_0000722, circ_0084765, circ_0002965, circ_0005311, circ_0001410, circ_0000043, circ_0001573, circ_0075796) could not even detect the differential expression of some circRNAs. (Chen JL, Rong N, Liu M, et al., The exosome-like vesicles derived from androgen exposed-prostate stromal cells promote epithelial cellsproliferation and epithelial-mesenchymal transition.Toxicol Appl Pharmacol,2021,411:115384.)
Further clinical validation failed to yield ideal results

Method used

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  • Application of exosomal circular RNA 0109315 as a target in the prevention and treatment of benign prostatic hyperplasia
  • Application of exosomal circular RNA 0109315 as a target in the prevention and treatment of benign prostatic hyperplasia
  • Application of exosomal circular RNA 0109315 as a target in the prevention and treatment of benign prostatic hyperplasia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Cigarette extract stimulates WPMY-1 cell proliferation.

[0026] (1) Human normal prostate stromal immortalized WPMY-1 cells were provided by the Chinese Academy of Sciences Stem Cell Bank.

[0027] (2) WPMY-1 cells were cultured with DMEM medium (containing 10 fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin).

[0028](3) Hongjinlong brand cigarettes (with nicotine, tar and carbon monoxide contents of 0.6, 8 and 11 mg / stick, respectively) were purchased from Wuhan Tobacco (Group) Co., Ltd.

[0029] (4) Fresh 10% cigarette smoke extract (Cigarette smoke, CS) was prepared by injecting smoke from a burning cigarette into 10 ml of culture medium and filtering it through a 0.22 μm filter membrane. 5% CS was obtained by dilution in medium.

[0030] (5) WPMY-1 cells were incubated with 5% CS for 48 hours.

[0031] (6) Collect 50 mL of cell supernatant, and extract exosomes with an exosome extraction kit (Guangzhou Jisai Biotechnology Co., Ltd.).

Embodiment 2

[0033] Whole transcriptome sequencing of WPMY-1 cell supernatant exosomes.

[0034] Collect WPMY-1 cells (control group) and exosomes from the cell supernatant after incubation with 5% CS in Example 1 for 48 hours, and each group has 3 biological repetitions.

[0035] The physiological conditions of WPMY-1 exosomes from human normal prostate stromal immortalized cells are as follows: figure 1 shown.

[0036] Extract the total RNA of exosomes, measure the RNA concentration with NanoDrop ND-1000 instrument, and the OD260 / OD280 value of 1.8-2.1 is qualified. High-throughput sequencing was provided by Shanghai Yunxu Biotechnology Co., Ltd. The sequencing library was constructed after removing ribosomal RNA from the sample. Quality control and quantification of the library were carried out by BioAnalyzer 2100system, and 150bp double-end sequencing was performed by illumina Hiseq4000 sequencer. STAR software (v2.5.1b) was used to align high-quality reads to the reference genome / ...

Embodiment 3

[0039] Application of hsa_circ_0109315 as a marker of benign prostatic hyperplasia:

[0040] Exosomal hsa_circ_0109315 levels in peripheral blood of healthy volunteers and patients with benign prostatic hyperplasia:

[0041] (1) Clinical specimens were collected from August 2020 to March 2021. Seven samples of peripheral blood were collected from BPH patients and healthy volunteers from Pu Ai Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology. The research involved in this application was approved by the Academic Ethics Review Committee of Jianghan University and the Ethics Committee of Puai Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.

[0042] (2) Extract exosomes with an exosome extraction kit (Guangzhou Jisai Biotechnology Co., Ltd.).

[0043] (3) hsa_circ_0109315 is 4296bp in length, and its corresponding parental gene is located at chr19:23541232-23545527;

[0044] (4) The seque...

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Abstract

The invention belongs to the field of biotechnology and medicine, and specifically relates to the application of exosome circular RNA 0109315 as a target in the prevention and treatment of benign prostatic hyperplasia. The applicant found that inhibiting hsa_circ_0109315 has a certain improvement effect on benign prostatic hyperplasia induced by various risk factors. The benign prostatic hyperplasia marker exosomal circular RNA hsa_circ_0109315 of the present invention has expression differences in the peripheral blood of healthy subjects and benign prostatic hyperplasia patients, so it can be used as a novel biomarker for the detection of benign prostatic hyperplasia. The present invention finds for the first time that exosome hsa_circ_0109315 can be used as a biomarker for the diagnosis of benign prostatic hyperplasia and a drug treatment target, which provides a new reference index for the development of drugs for the prevention and treatment of benign prostatic hyperplasia.

Description

technical field [0001] The invention belongs to the field of biotechnology and benign prostatic hyperplasia, and specifically relates to the application of exosomal circular RNA hsa_circ_0109315 as a target in the prevention and treatment of benign prostatic hyperplasia. Background technique [0002] Benign prostatic hyperplasia (BPH) is a common disease in middle-aged and elderly men, and its incidence is on the rise and increases with age. According to reports, the incidence of BPH in men over 50, 60 and 80 years old is about 25%, 50% and 83% respectively (WuF, Ding S, Li X, et al. Elevated expression of HIF-1α inactively growing prostate tissuesis associated with clinical features of benign prostatic hyperplasia. Oncotarget, 2016, 7(11): 12053-62.). In addition, BPH is also the most common cause of lower urinary tract symptoms including dysuria and urinary storage disorders in men (Gratzke C, Bachmann A, Descazeaud A, et al.EAU guidelines on the assessment of non-neuroge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883A61K45/00A61K31/7088A61P13/08
CPCC12Q1/6883A61K45/00A61K31/7088A61P13/08C12Q2600/158C12Q2600/178C12Q2600/136
Inventor 陈镜楼刘敏刘建华容楠
Owner JIANGHAN UNIVERSITY
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