Application of CDCA8 in preparation of drug for treating ovarian cancer

A technology for ovarian cancer and ovarian cancer cells, applied in the field of biomedicine, can solve the problem of no research on the relationship between chemotherapy and drug resistance of ovarian cancer, and achieve the effect of inhibiting cell apoptosis

Active Publication Date: 2020-12-01
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no research on the relationship between CDCA8 and t...

Method used

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  • Application of CDCA8 in preparation of drug for treating ovarian cancer
  • Application of CDCA8 in preparation of drug for treating ovarian cancer
  • Application of CDCA8 in preparation of drug for treating ovarian cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Detection of CDCA8 expression level in fallopian tube umbrella and ovarian cancer tissues:

[0039] The expression level of CDCA8 was detected from two levels of mRNA and protein. First, use TRIzol (15596018, Invitrogen) to extract RNA and reverse transcribe it into cDNA, and continue to use RT-qPCR to detect CDCA8 mRNA; protein level detection includes Western blot and immunohistochemical methods. Western blot was established on the basis of lysing tissue with IP cell lysate (P0013, Biyuntian), centrifuging and measuring the total protein concentration, and using Western Blot method to detect the expression level of CDCA8 protein. In addition, the fallopian tube umbrella tissue and ovarian cancer tissue collected by Qilu Hospital of Shandong University were selected, and the CDCA8 protein level was detected by immunohistochemical method.

[0040] Such as figure 1 As shown in A and 1B, the expression of CDCA8 mRNA and protein in ovarian cancer tissues is higher than t...

Embodiment 2

[0042] CDCA8 promotes ovarian cancer proliferation experiment:

[0043] A2780, SKOV3 and HEY cells were transfected with NC, CDCA8 siRNA, overexpression control (PCMV), and CDCA8 overexpression plasmid (CDCA8) for 48 hours;

[0044] CDCA8 knockdown and overexpression efficiencies were detected by Western Blot. The effects of CDCA8 expression on the proliferation of ovarian cancer cell lines were detected by MTT and plate cloning assays; the in vivo tumor-bearing experiments were used to verify the effect of CDCA8 on tumorigenesis in vivo.

[0045] MTT: A2780, SKOV3 and HEY cells were transfected with NC, CDCA8 siRNA, PCMV, and CDCA8 for 24 hours, digested, centrifuged, counted, and seeded in a 96-well plate with 600-1000 cells per well and 6 secondary wells for each group. Placed in an incubator to continue culturing for 1-5 days, and the cell viability was detected by the MTT (Sigma) method.

[0046] Plate cloning: A2780, SKOV3, and HEY cells were transfected with NC, CDCA8...

Embodiment 3

[0050] CDCA8 promotes invasion and migration of ovarian cancer cells:

[0051] Transwell experiment: A2780, SKOV3 and HEY cells were transfected with NC, CDCA8 siRNA, PCMV, CDCA8 24 hours after digestion, centrifugation and counting, 1×10 5 ~2×10 5 Each cell was resuspended in 200 μL of serum-free medium and placed on the upper layer of the chamber, and 700 μL of medium containing 20% ​​serum was placed on the lower layer. After culturing for a certain period of time, the effect of CDCA8 on cell invasion and migration was detected.

[0052] Such as image 3 As shown in A, interference with CDCA8 expression inhibits the invasion ability of ovarian cancer cell lines, and overexpression of CDCA8 promotes the invasion ability of ovarian cancer cells; image 3 As shown in B, interfering with the expression of CDCA8 inhibits the migration ability of ovarian cancer cell lines, and overexpressing CDCA8 promotes the migration ability of ovarian cancer cells.

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Abstract

The invention belongs to the field of biological medicines, and particularly relates to an application of CDCA8 in the preparation of a drug for treating ovarian cancer. The invention provides an application of a reagent for inhibiting CDCA8 gene expression or inhibiting a CDCA8 gene expression product in the preparation of a drug for treating ovarian cancer and/or inhibiting proliferation, invasion and migration of ovarian cancer cells and at the same time promoting ovarian cancer cell cycle arrest and apoptosis. In addition, the inhibition of the expression of CDCA8 or the inhibition of theCDCA8 gene expression product can enhance the sensitivity of ovarian cancer cells to chemotherapeutic drugs cisplatin and olaparib by adjusting the cell cycle and inhibiting DNA damage repair. A new thought is provided for screening drugs for treating ovarian cancer, especially serous ovarian cancer, and a new method is provided for enhancing the sensitivity to traditional ovarian cancer chemotherapeutic drugs.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of CDCA8 in the preparation of drugs for treating ovarian cancer. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Ovarian cancer is one of the most common gynecological malignancies, and its mortality rate ranks first among gynecological malignancies. Serous ovarian carcinoma (SOC) is the most common epithelial ovarian cancer, with high malignancy and poor prognosis. [0004] At present, the first-line treatment for ovarian cancer is still cytoreductive surgery and platinum / paclitaxel-based chemotherapy, but most patients will relapse and drug resistance. ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P35/00A61P35/04A61K33/243A61K31/502
CPCA61K45/00A61P35/00A61P35/04A61K33/243A61K31/502A61K2300/00
Inventor 孔北华齐贡花马汉林苑存忠
Owner SHANDONG UNIV QILU HOSPITAL
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