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Application of STAT1 serving as ovarian cancer treatment target point

A technology for ovarian cancer and ovarian cancer cells, applied in the field of biomedicine, to achieve the effect of strong clinical application possibility

Active Publication Date: 2017-01-18
JINSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, how STAT1 signaling pathway and TGF-β1 signaling pathway affect the occurrence and development of ovarian cancer in ovarian cancer, and the interaction between them has not been reported

Method used

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  • Application of STAT1 serving as ovarian cancer treatment target point
  • Application of STAT1 serving as ovarian cancer treatment target point
  • Application of STAT1 serving as ovarian cancer treatment target point

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 STAT1 and TGF-β receptor combine to form a complex

[0019] STAT1α-myc plasmid: the target fragment (GenBank accession#NM_007315) was inserted between the Kpn I and Sac II sites of pcDNA4 / TO / myc-His(B) vector (Invitrogen, Catalog no.V1030-20).

[0020] STAT1β-myc plasmid: the target fragment (GenBank accession#NM_139266) was inserted between the pcDNA4Kpn I and Sac II sites.

[0021] ALK1-HA, ALK5-HA, TβRII-HA plasmids: construction reference Xu G, Barrios-Rodiles M, Jerkic M, Turinsky AL, Nadon R, Vera S, et al. Novel protein interactions with endoglin and activin receptor-like kinase 1: potential role in vascular networks. Mol Cell Proteomics 2014,13(2):489-502.

[0022] STAT1α-myc and ALK1-HA, ALK5-HA or TβRII-HA plasmids were co-transfected into 293T cells. The plasmid transfection kit was DNA18 Transfection Reagent (GBC lifetech, Miami, FL, USA), and the co-transfection concentration was 2 μg / ml, 293T cells were cultured in DMEM medium supplemented wit...

Embodiment 2

[0023] Example 2 Effect of STAT1 on proliferation of ovarian cancer cell lines

[0024] In the overexpression experiment ( figure 2 A), the ovarian cancer cells OVCAR-3 and SK-OV-3 were transfected with vector pcDNA4 and plasmid pSTAT1α-myc or pSTAT1β-myc, and the plasmid transfection kit was DNA Transfection Reagent (GBC lifetech, Miami, FL, USA), The co-transfection concentration was 2 μg / ml, and the siRNA transfection kit was tremeGENE siRNAtransfection reagent (Roche Diagnostics); OVCAR-3 was cultured in RPMI-1640 medium, supplemented with 10% serum. SK-OV-3 was cultured in DMEM medium supplemented with 10% serum. Cell Proliferation Reagent (WST-1 kit, Roche) was used to detect cell proliferation by WST-1 method, and the absorbance at 450 nm was detected 48 hours after transfection with a microplate reader. In siRNA interference experiments ( figure 2 B), the STAT1-siR sequence number used is GenBank accession#, NM_007315, the STAT1-siR sequence is: Sense: 5'-GCGUAAUC...

Embodiment 3

[0025] Example 3 Effect of STAT1 on the Migration Ability of Ovarian Cancer Cells (Cell Scratch Test)

[0026]Use a marker pen to evenly draw a horizontal line on the back of the 6-well plate, plate SK-OV-3 cells, use a 200μl pipette tip to scratch perpendicular to the horizontal line on the back the next day, wash off the scratched cells with PBS, and add serum-free culture base. Plasmid transfection or siRNA transfection of cells, the transfection conditions were the same as in Example 2. Fresh medium was changed after 4-6h, and photographs were taken at 0h, 24h, 48h, and 72h. image 3 A and image 3 C is the pictures taken at each time point after the scratch, image 3 B and image 3 D is to measure the average distance of the three scratches and make a graph. The distance between the scratches in the STAT1α group or the STAT1β group is larger than that in the pcDNA4 group at each time point, indicating that the migration ability of the cells in the STAT1α group or the S...

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Abstract

The invention relates to application of STAT1 serving as ovarian cancer treatment target point. Experiments prove that the STAT1 directly combines with a TGF-beta receptor (ALK1, ALK5 and TbetaRII), proliferation, migration and invasion of ovarian cancer cells can be promoted after STAT1overexpression, and proliferation, migration and invasion of the ovarian cancer cells can be inhibited after STAT1 knocking-down. Therefore, STAT1 reduction processing is conducted on patients with high ovarian cancer STAT1 expressions, a potential treatment effect exists. An STAT1 signal path mutually acts with a TGF-beta1 signal path to influence the proliferation, migration and invasion capability of ovarian cancers. The STAT1 is a potential novel ovarian cancer treatment target point and has a great clinical application value.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to the use of STAT1 as a therapeutic target for ovarian cancer. Background technique [0002] Ovarian cancer is one of the common malignant tumors of female reproductive organs. Due to the complicated embryonic development, tissue anatomy and endocrine function of the ovary, the early symptoms are not obvious, and it is quite difficult to distinguish the tissue type and benign from malignant ovarian tumors before surgery. . The etiology of ovarian cancer is not clear, and it may be related to the following aspects: external factors of cancer (including chemical, physical, biological and other carcinogenic factors); internal factors of cancer (including immune function, endocrine, genetics, age, mental factors, etc.) , as well as dietary malnutrition and bad living habits. Mostly occurs in perimenopausal women. Those over 35 years old are mostly epithelial ovarian canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7105A61K48/00A61P35/00
CPCA61K31/7105A61K45/00
Inventor 许国雄田晓玲管文彩
Owner JINSHAN HOSPITAL FUDAN UNIV
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