ZNF695 as marker and treatment target of prostate cancer bone metastasis

A ZNF695, prostate cancer technology, applied in the field of biomedicine, can solve the problems of reducing the quality of life and life expectancy of PCa patients, and clinical treatment difficulties

Pending Publication Date: 2020-08-18
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

Therefore, once bone metastasis occurs, it not only seriously reduces the quality of life and life expectancy of PCa patients, but also brings great difficulties to clinical treatment.

Method used

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  • ZNF695 as marker and treatment target of prostate cancer bone metastasis
  • ZNF695 as marker and treatment target of prostate cancer bone metastasis
  • ZNF695 as marker and treatment target of prostate cancer bone metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1, mRNA and protein detection of ZNF695

[0035] 1.1 ZNF695 mRNA detection

[0036] Real-time quantitative PCR method was used: ① RNA was extracted using Trizol reagent, and 2 μg of RNA was subjected to reverse transcription reaction. After the reaction was completed, the cDNA was stored at -20°C for later use. ② Prepare the SYBRGreen mix (Roche) required for DNA amplification according to the instructions of the detection kit, as well as ZNF695 detection primers and internal reference primers. The design and synthesis of the primers are from Guangzhou Ruibo Biotechnology Co., Ltd. In the Bio-rad real-time quantitative PCR instrument, the amplification reaction conditions were set according to the requirements of the kit instructions, and 40 cycles of reactions were performed. The GAPDH gene was used as an internal reference, and at least 3 auxiliary wells were set for all samples. ③ The relative expression of target gene DNA passed 2 -ΔΔCt calculated by the ...

Embodiment 2

[0043] Example 2. Low expression of ZNF695 inhibits bone metastasis ability of prostate cancer cells in vivo

[0044] 2.1 In vitro culture of PCa bone metastasis cells

[0045] PCa bone metastasis PC-3 cell line was cultured in CO 2 The concentration is 5%, and the temperature is in an incubator at 37°C. Cells were grown in Ham's F-12 medium (10% fetal bovine serum). Observe the cell state and cell density, and replace the medium regularly (generally about 2-3 days), and it is suitable for experiment use when the cell density reaches 80%.

[0046] 2.2 Transfection

[0047] The RNA interference plasmids (siZNF695#1, siZNF695#2) and their control plasmids (vector) used to down-regulate the expression of ZNF695 were designed and synthesized by Guangzhou Ruibo Biotechnology Co., Ltd. Before transfection, prostate cancer PC-3 cells were incubated with 10 5 Each well was inoculated on a culture plate, and diluted RNA interference plasmids, control plasmids and liposomes were ad...

Embodiment 3

[0054] Example 3. Low expression of ZNF695 inhibits bone metastasis of prostate cancer cells in vitro

[0055] 3.1 In vitro culture of PCa cells

[0056] PC-3 cell line and VcaP cell line were cultured in CO 2 The concentration is 5%, and the temperature is in an incubator at 37°C. Cells were grown in Ham's F-12 medium (10% fetal bovine serum). Observe the cell state and cell density, and replace the medium regularly (generally about 2-3 days), and it is suitable for experiment use when the cell density reaches 80%.

[0057] 3.2 Transfection

[0058] In the same way as in 2.2, siZNF695#1 and its control plasmid were transfected into prostate cancer PC-3 cells or VcaP cells, respectively, to obtain prostate cancer cells with low expression of ZNF695.

[0059] 3.3 Migration and invasion experiments

[0060] ①Use 50mg / L matrigel to coat or not coat the polycarbonate filter membrane of the transwell chamber. ② Take 100 μL of serum-free medium containing about 5×104 PCa cells...

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Abstract

The invention provides ZNF695 as a marker and a treatment target of prostate cancer bone metastasis. It is determined that the ZNF695 is highly expressed in a prostate cancer tissue and is particularly specific in the prostate cancer tissue accompanied by bone metastasis, and in-vivo prostate cancer bone metastasis can be remarkably inhibited by down-regulation of the ZNF695. Therefore, the ZNF695can be used as an early diagnosis marker of prostate cancer and bone metastasis thereof and a treatment target of prostate cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and more specifically relates to ZNF695 as a marker and therapeutic target of prostate cancer bone metastasis. Background technique [0002] Most cancer patients die not from the original cancer itself, but from metastatic cancer. Tumor metastasis is the fundamental reason for the failure of clinical treatment of malignant tumors. Bone is one of the most common distant metastatic organs of solid tumors. A foreign autopsy study of patients who died of prostate cancer (PCa) showed that about 80% of the cases had bone metastases. In my country, with the change of nutritional structure and the trend of aging, the incidence and mortality of PCa have a significant increase trend. At present, there is no definite molecular mechanism to elucidate PCa bone metastasis, and no related target can effectively predict PCa bone metastasis. Therefore, once bone metastasis occurs, it not only seriously reduces the q...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574
CPCC12Q1/6886G01N33/57434C12Q2600/158C12Q2600/112
Inventor 黄帅瓦庆德黄胜唐欲博梁宝霞
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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