Application of reagent for down-regulating gene expression in preparation of prostatic cancer drugs

A prostate cancer and prostate technology, applied in the field of tumor biology, can solve the problems of incompletely clear pathogenesis of prostate cancer and complex pathogenesis of prostate cancer, and achieve the effect of inhibiting proliferation and metastasis

Active Publication Date: 2021-09-07
SHANDONG PROVINCIAL HOSPITAL AFFILIATED TO SHANDONG FIRST MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pathogenesis of prostate cancer is very complex, so the pathogenesis of the occurrence and progression of prostate cancer is not yet fully understood.

Method used

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  • Application of reagent for down-regulating gene expression in preparation of prostatic cancer drugs
  • Application of reagent for down-regulating gene expression in preparation of prostatic cancer drugs
  • Application of reagent for down-regulating gene expression in preparation of prostatic cancer drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Tissue total RNA extraction

[0041] (1) Before starting the experiment, soak the equipment required for the experiment in 0.1% DEPC water overnight, and dry it in the oven for later use;

[0042] (2) Take about 50 mg of fresh tumor tissue and paracancerous tissue, use ophthalmic scissors to cut the tissue into a pre-cooled mortar, pour liquid nitrogen into it, grind the tissue quickly, and transfer the tissue to In a new RNase-free 1.5ml EP tube;

[0043] (3) Add 1ml of Trizol lysate, let stand at room temperature for 5min, add 200ul of chloroform, vortex for 30s with a vortexer, and place on ice for 10min;

[0044] (4) Put the EP tube into a centrifuge at 4°C and centrifuge at 12000g for 15 minutes. After the end, carefully transfer the supernatant to a new 1.5ml EP tube;

[0045] (5) Add an equal volume of isopropanol and mix well, place on ice for 10 minutes, put the EP tube into a centrifuge at 4°C, centrifuge at 12000g for 10 minutes, and discard the supernatant...

Embodiment 2

[0068] Detection of the expression difference of Lnc-RP5-952N6.1 in prostate cancer cells

[0069] (1) Extract RNA from BPH-1 cells, DU-145 cells, LnCap cells, PC3 cells, and VCaP cells according to the method in Example 1;

[0070] (2) According to the method of Example 1, the expression difference of Lnc-RP5-952N6.1 in BPH-1 cells, DU-145 cells, LnCap cells, PC3 cells, and VCaP cells was detected.

[0071] Among them, the relative expression level of Lnc-RP5-952N6.1 in DU-145 cells was 3.606±0.243;

[0072] The relative expression level of Lnc-RP5-952N6.1 in LnCap cells was 2.613±0.199;

[0073] The relative expression level of Lnc-RP5-952N6.1 in PC3 cells was 2.332±0.452;

[0074] The relative expression level of Lnc-RP5-952N6.1 in VCaP cells was 3.023±0.310;

[0075] The above results indicated that the relative expression of Lnc-RP5-952N6.1 in prostate cancer cells was significantly increased compared with BPH-1 in human prostatic hyperplasia cells, and the expression ...

Embodiment 3

[0077] Design shRNA sh-Lnc-RP5-952N6.1 for Lnc-RP5-952N6.1

[0078] (1) Design shRNA according to the sequence SEQ ID NO.5 of Lnc-RP5-952N6.1, and construct it on the pENTR™ / U6 plasmid, as follows:

[0079] Top Strand:

[0080] 5'-CACCGCGGCTTCCTCTAGAATAAAGCGAACTTTATTCTAGAGGAAGCCGC-3'; SEQ ID NO.6;

[0081] Bottom Strand

[0082] 5'-AAAAGCGGCTTCCTCTAGAATAAAGTTCGCTTTATTCTAGAGGAAGCCGC-3'; SEQ ID NO.7;

[0083] (2) Transfect sh-Lnc-RP5-952N6.1 into DU-145 cells, and detect the inhibitory effect of sh-Lnc-RP5-952N6.1.

[0084] Among them, in DU-145 cells transfected with sh-Lnc-RP5-952N6.1, the relative expression level of Lnc-RP5-952N6.1 was 0.198±0.031, indicating that sh-Lnc-RP5-952N6.1 can effectively inhibit Expression of Lnc-RP5-952N6.1.

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Abstract

The invention provides application of a reagent for down-regulating gene expression in preparation of prostatic cancer drugs, and belongs to the technical field of tumor biology. The invention finds that down regulating of high expression of the gene Lnc-RP5-952N6.1 in prostate cancer cells can effectively inhibit proliferation and metastasis of the prostate cancer cells, so that a reagent for down-regulating expression of the gene Lnc-RP5-952N6.1 can be used for preparing drugs for treating prostate cancer.

Description

[0001] This case is a divisional application. The invention name of the original application is: a prostate cancer marker and its therapeutic drug. The filing date of the original application is: 2021-02-03, and the application number of the original application is: 202110151318X. technical field [0002] The invention belongs to the technical field of tumor biology, and particularly relates to the application of a reagent for down-regulating gene expression in the preparation of prostate cancer medicine. Background technique [0003] Prostate cancer is a common urinary system malignancy in elderly men. With the change of diet and lifestyle and the aging of the population, the number of prostate cancer patients is increasing, which seriously threatens the health of middle-aged and elderly men. At the same time, prostate cancer is also showing a younger trend. The early clinical symptoms of prostate cancer patients are not obvious, and the patients are often already at an ad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61P35/00A61P35/04C12Q1/6886
CPCC12Q1/6886C12Q2600/178C12Q2600/158C12Q2600/136A61K45/00A61K31/713A61P35/00A61P35/04
Inventor 夏志明侯东省张亚洲周霞
Owner SHANDONG PROVINCIAL HOSPITAL AFFILIATED TO SHANDONG FIRST MEDICAL UNIVERSITY
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