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Application of miR-19a-3p in preparing diagnostic reagent and therapeutic drug for prostate cancer with osseous metastasis

1. The technology of mir-19a-3p and prostate cancer, which is applied in the field of molecular biology, can solve the problems that affect the treatment effect of tumors and have not yet

Active Publication Date: 2017-12-19
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chemoradiotherapy drugs not only act on the tumor, but can also act on the healthy tissues around the tumor. Therefore, while killing the tumor, they also bring great side effects to the body, which ultimately affects the therapeutic effect on the tumor.
There is no report on the relationship between miR-19a-3p and prostate cancer bone metastasis

Method used

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  • Application of miR-19a-3p in preparing diagnostic reagent and therapeutic drug for prostate cancer with osseous metastasis
  • Application of miR-19a-3p in preparing diagnostic reagent and therapeutic drug for prostate cancer with osseous metastasis
  • Application of miR-19a-3p in preparing diagnostic reagent and therapeutic drug for prostate cancer with osseous metastasis

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0024] Experimental Example 1. Low expression of miR-19a-3p in prostate cancer tissue with bone metastases

[0025] method:

[0026] RNA extraction, reverse transcription and real-time quantitative RT-PCR detection of miR-19a-3p: TRIzol lysate (Life Technologies) was used to extract total RNA from all tissues and cells, and reverse transcription was performed according to the instructions of Revert Aid First StrandcDNA SynthesisKit (Thermo Fisher) , using SYBR Green I reagent and ABI7500HT real-time quantitative PCR system (Biosystems), using 2-ddCt relative quantitative method for fluorescence quantitative detection, using U6 as an internal reference, to calculate the relative expression of miR-19a-3p, miR-19a- The primers for 3p and U6 were designed, synthesized and purified by Guangzhou Ruibo Biotechnology Co., Ltd. The sequence of miR-19a-3p refers to miRBase number MIMAT0000073.

[0027] result:

[0028] The results of miR-19a-3p expression in clinical specimens and cel...

experiment example 2

[0029] Experimental example 2. Overexpression of miR-19a-3p can inhibit the migration and invasion of prostate cancer in vitro

[0030] method:

[0031]The human miR-19a-3p (miRBase number MIMAT0000073) gene was cloned into the pMSCV-puro retroviral vector (Clontech Company) to obtain the miR-19a-3p expression plasmid, and the antisense nucleotide of human miR-19a-3p ( The anti-miR-19a-3p) sequence was cloned into the pH1 plasmid, which was synthesized by Shanghai Jikai Gene Chemical Technology Co., Ltd., and the miR-19a-3p expression plasmid, anti-miR-19a-3p expression plasmid, and the corresponding The negative control plasmid was transfected into PC-3 cells and C4-2B cells, the cell migration and invasion were detected by Transwell chamber method, and the expression of miR-133b was detected by fluorescence quantification

[0032] result:

[0033] The constructed expression plasmid was transfected into PC-3 cells and C4-2B cells, and the expression level of miR-19a-3p was ...

experiment example 3

[0035] Experimental Example 3. Overexpression of miR-19a-3p can inhibit bone metastasis of prostate cancer in vivo

[0036] Experimental grouping and animal model construction:

[0037] ① miR-19a-3p group

[0038] The miR-19a-3p constructed in Experimental Example 2 was transfected into PC-3- cells to form PC-3 cells stably and highly expressing miR-19a-3p, which were injected into six 3- to 4-week-old rats through the tibia Right tibia of male SCID mice.

[0039] ② Control group

[0040] The negative control plasmid pMSCV-puro of Experimental Example 2 was transfected into PC-3 cells to form plasmid-controlled PC-3 cells, which were injected into the left side of six 3- to 4-week-old male SCID mice through the tibia. in the tibia.

[0041] Imaging detection:

[0042] X-ray films were taken to check the bone destruction at the bone metastases, and the scores were scored according to the degree of bone destruction, 0: no bone destruction, 1: slight bone destruction; 2: a s...

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Abstract

The invention discloses an application of miR-19a-3p in preparing a diagnostic reagent and a therapeutic drug for prostate cancer with osseous metastasis. The invention researches and finds that the miR-19a-3p has low expression in prostate cancer with osseous metastasis, the miR-19a-3p low expression is related to the prostate cancer with osseous metastasis, the over-expression miR-19a-3p is capable of in vitro restraining migration and invasion of prostate cancer with osseous metastasis and in vivo restraining osseous metastasis of prostate cancer, so that the miR-19a-3p can be used for preparing diagnostic reagent and the therapeutic drug for prostate cancer with osseous metastasis.

Description

technical field [0001] The invention belongs to the field of molecular biology, and more specifically relates to the application of miR-19a-3p in the preparation of diagnostic reagents and therapeutic drugs for prostate cancer bone metastasis. Background technique [0002] Prostate cancer (PCa) is one of the most common malignant tumors of the male genitourinary system. Cancer metastasis will greatly increase the mortality of cancer patients. Bone is the second most common site of cancer metastasis. Common cancers with bone metastasis include breast cancer, prostate cancer, lung cancer and kidney cancer. In my country, more than 80% of prostate cancer patients have developed bone metastases at the time of diagnosis, and bone metastasis is an important cause of death in prostate cancer patients. Therefore, early diagnosis of bone metastases is the key to clinical diagnosis, treatment and prognosis. In addition, for patients with advanced prostate cancer, the effect of surgic...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K48/00A61K31/7088A61P35/04
CPCA61K31/7088C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 黄帅瓦庆德唐欲博郭远清彭新生
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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