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Marker LncRNA ZNF518A for diagnosing castration-resistant prostate cancer and/or performing prognosis assessment, and application of marker LncRNA ZNF518A

A technology for prostate cancer and castration resistance, applied in the field of molecular biology, to achieve strong specificity, practical clinical value, and the effect of promoting occurrence and development

Active Publication Date: 2020-06-05
SHANGHAI TONGJI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is a detection method that cannot be replaced by other methods at present, and there has been no relevant report so far

Method used

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  • Marker LncRNA ZNF518A for diagnosing castration-resistant prostate cancer and/or performing prognosis assessment, and application of marker LncRNA ZNF518A
  • Marker LncRNA ZNF518A for diagnosing castration-resistant prostate cancer and/or performing prognosis assessment, and application of marker LncRNA ZNF518A
  • Marker LncRNA ZNF518A for diagnosing castration-resistant prostate cancer and/or performing prognosis assessment, and application of marker LncRNA ZNF518A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Immunofluorescence detection of AR-V7

[0052] 1 Experimental method

[0053] Immunohistochemical detection (Immunohistochemistry): Specimens were taken from tumor tissue and adjacent normal prostate tissue, dehydrated and embedded in paraffin, each case was sliced ​​into 6 pieces of paraffin block, the tissue thickness was 4um, and the slices were baked in a 70°C incubator for 15 minutes , deparaffinized, rinsed with PBS 3 times, 2 min each time. The specimens were placed in an antigen retrieval box, soaked in 0.01mol citrate buffer, heated in a microwave oven for 15 minutes, and cooled to room temperature. Rinse with PBS 3 times, 2 min each time. Add 50ul of peroxidase blocking agent dropwise to the slices, incubate at room temperature in the dark for 15min, wash with PBS 3 times, 2min each time. Remove the PBS solution, add goat serum dropwise to the slices, incubate at room temperature in the dark for 20 minutes, remove excess serum without washing, add...

Embodiment 2

[0056] Example 2 Verifying the relationship between LncRNA ZNF518A and prostate cancer

[0057] 1 Experimental method

[0058] 15 samples of benign prostatic hyperplasia and 30 samples of prostate cancer were selected, among which PSA>714 cases and PSA<716 cases, the RNA was extracted, reversed to cDNA, quantified, added 10ul of cDNA template and 0.5U of Taq enzyme, 97℃5 minutes, immediately ice-water bath, mix well, 95°C for 5 minutes, 94°C for 30 seconds for denaturation, 40 seconds for annealing, 72°C for 30 seconds for extension, 72°C for 7 minutes for terminal extension for 28-36 cycles, and 4°C for incubation.

[0059] 2 Experimental results

[0060] see results figure 1 As shown, LncRNA ZNF518A was significantly increased in prostate cancer, and the higher the degree of malignancy, the higher its expression.

Embodiment 3

[0062] 1 Experimental method

[0063] 1.1 Select C4-2 enzalutamide-resistant and 22RV-1 docetaxel-resistant cells, one group knocks out LncRNAznf518A, and the other group serves as a control; add 500ul TRIzol reagent to the 6well culture plate covered with two kinds of cells, Place on ice for 5 minutes, pipette evenly, transfer to EP tube; add 100ul of chloroform, place at room temperature for 5 minutes, centrifuge at 12000g at 4°C for 10 minutes, transfer 400ul of the upper aqueous phase to another EP tube, add 250ul of isopropanol, mix at room temperature Stand for 10 minutes, centrifuge at 12000g at 4°C for 10 minutes, discard the supernatant, add 1ml of 75% ethanol, centrifuge at 7500g at 4°C for 5 minutes, discard the supernatant, air dry for 5-10 minutes, dissolve in DEPC water to 30ul, quantify, add cDNA Template 10ul and Taq enzyme 0.5U, 97°C for 5 minutes, immediately ice-water bath, mix well, 95°C for 5 minutes, 94°C for 30 seconds for denaturation, 40 seconds for an...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a marker LncRNA ZNF518A for diagnosing castration-resistant prostate cancer and / or performing prognosis assessment,and an application of the marker LncRNA ZNF518A. The experimental result indicates that the LncRNA ZNF518A has binding sites of miR-101-3P and AR, the miR-101-3P can also be bound with AR-V7 3-'UTR, and the result is verified by a luciferase report gene, so that the inventor finds that the LncRNA ZNF518A can adjust and control the expression of AR-V7 through competitive binding with miR-101-3P, and further generation and development of the CRPC are promoted. The marker LncRNA ZNF518A has the advantages that through double detection for the expression of the LncRNA ZNF518A and AR-V7 fluorescence immunochemistry detection, feasibility and credibility of clinical application are improved, the marker LncRNA ZNF518A can be directly applied to clinical application, and the characteristic of themarker LncRNA ZNF518A is incomparable for other molecular biology detection methods at present.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular, it relates to castration-resistant prostate cancer diagnosis or prognosis evaluation marker LncRNA ZNF518A and its application. Background technique [0002] Prostate cancer (PCa) is one of the most common and deadly malignant tumors in European and American men (Siegel RL, Miller KD, Jemal A. Cancer statistics, 2015. CA Cancer J Clin 2015; 65(1): 5- 29). In big cities such as Beijing and Shanghai in my country, prostate cancer has become the most common tumor of the male genitourinary system, and most patients are diagnosed at an advanced stage (Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, et al. .Cancer statistics in China,2015.CA Cancer J Clin 2016,66(2):115-32, Ye Dingwei, Zhu Yao. Epidemiological overview and enlightenment of prostate cancer in China. Chinese Journal of Surgery 2015;53(4):249 -52). For advanced prostate cancer, anti-androgenic therapy is comm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00C12N15/11
CPCC12Q1/6886A61K45/00A61P35/00C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 吴刚王新安周娟徐成党姚怿聪施燕卞崔冬吴登龙
Owner SHANGHAI TONGJI HOSPITAL
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