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Application of blocking antibody T5E3 to reversing of tumor multidrug resistance

A multi-drug resistance and antibody technology, applied in the direction of anti-tumor drugs, antibodies, anti-animal/human immunoglobulin, etc., to achieve good development prospects and non-toxic side effects

Active Publication Date: 2014-04-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] At present, there is no application of TRPC5 blocking antibody T5E3 in reversing tumor multidrug resistance at home and abroad

Method used

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  • Application of blocking antibody T5E3 to reversing of tumor multidrug resistance
  • Application of blocking antibody T5E3 to reversing of tumor multidrug resistance
  • Application of blocking antibody T5E3 to reversing of tumor multidrug resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Down-regulation of P-gp expression in multidrug-resistant tumor cells by TRPC5 blocking antibody T5E3

[0027] experimental method:

[0028] MCF-7 / ADM was inoculated on a six-well plate at a density of 300,000 cells per well. After culturing for 12 hours, different concentrations of T5E3 (10 μg / mL, 20 μg / mL) were added to each well, and serum IgG from New Zealand rabbits before immunization was used.

[0029] As the control group (Ctl), the cells were cultured routinely, and the cells in each well were collected after 24 hours. According to the conventional operation method, Western blot was used to detect the expression of P-gp before and after TRPC5 was inhibited.

[0030] Tubulin was used as an internal reference, the primary antibody for incubation with βtubulin was βtubulin antibody (SC-9104), and the secondary antibody for incubation was horseradish peroxidase-labeled goat anti-rabbit IgG (H+L).

[0031] Experimental results:

[0032] See the experime...

Embodiment 2

[0034] Example 2 Reversal effect of TRPC5 blocking antibody T5E3 on multidrug resistance of tumor cells in vitro

[0035] experimental method:

[0036] MCF-7 / ADM and MCF-7 / WT were inoculated on 25cm2 When the MCF-7 / ADM confluence was 80%, TRPC5 blocking antibody T5E5 was added at 20 μg / mL, and New Zealand rabbit pre-immune serum IgG was used as the control group, and the cells were routinely cultured, and after 24 hours Collect the above-treated MCF-7 / ADM and untreated MCF-7 / WT, inoculate in a 96-well plate at a density of 7000 cells per well, add doxorubicin according to the concentration gradient after 24 hours, and culture for 48 hours Afterwards, the drug sensitivity of the cells was detected by MTT assay.

[0037] Experimental results:

[0038] See the experimental results figure 2 . Depend on figure 2 It can be seen that the sensitivity of multidrug-resistant tumor cells (MCF-7 / ADM) to doxorubicin was significantly lower than that of wild-type tumor cells (MCF-7 / W...

Embodiment 3

[0039] Example 3 Reversal effect of TRPC5 blocking antibody T5E3 on multidrug resistance of tumor cells in vivo

[0040] experimental method:

[0041] The TRPC5 blocking antibody T5E3 was injected every three days at the tumor site of the drug-resistant tumor-bearing mice, and divided into two experimental groups: the middle and low dose group (2 μg), and the high dose group (4 μg). New Zealand rabbits were immunized Pre-serum IgG was used as the control group; ADM (4 mg / kg mouse weight) was continued to be injected on the second day after each injection of T5E3, and the tumor was removed at different time periods, and the tumor volume was taken and measured for a total of 30 days.

[0042] Experimental results:

[0043] See the experimental results image 3 . Depend on image 3 It can be seen that the TRPC5 blocking antibody T5E3 can significantly inhibit the growth of drug-resistant tumors in a dose-dependent manner. The experimental results show that T5E3 has an obviou...

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Abstract

The invention provides the novel application of the blocking antibody T5E3 of the TRPC5, particularly the application of the blocking antibody T5E3 of the TRPC5 to the reversing of tumor multidrug resistance. The blocking antibody is prepared by using the E3 peptide fragment which is obtained by artificial synthesis or gene expression as an antigen to immune an animal. The tumor multidrug resistance is generated by the mediation of P-gp (p-glycoprotein). The invention is characterized in that: the close relation of the TRPC5 and the tumor multidrug resistance is found, and the expression level of the P-gp for the tumor to generate the multidrug resistance in the multidrug resistance tumor cell can be indirectly increased and mediated. The blocking antibody T5E3 of the TRPC5 has the function of reversing the multidrug resistance of the tumor cells in vivo and vitro significantly and can not cause toxic and side effects to the experimental animal. The invention provides a novel idea for reversing the tumor multidrug resistance. The blocking antibody T5E3 is a tumor multidrug resistance reversing agent with good development prospect.

Description

technical field [0001] The present invention relates to a new application of the TRPC5 blocking antibody T5E3, in particular to the application of the TRPC5 blocking antibody T5E3 in reversing tumor multidrug resistance. Background technique [0002] Tumor multidrug resistance (MDR) refers to the generation of cross-resistance to anti-tumor drugs with different structures and mechanisms of action while tumor cells develop resistance to one anti-tumor drug, thereby greatly reducing the efficacy of anti-tumor drugs. curative effect. Transmembrane protein-mediated MDR is the most classic mechanism of MDR production, the most common of which is P-glycoprotein. P-glycoprotein (P-glycoprotein, P-gp), also known as p170 glycoprotein, is an ATP-dependent membrane transport protein encoded by the multidrug resistance gene MDR1. P-gp is a drug pump that can A variety of drugs are pumped out of the cells, reducing the accumulation of intracellular drugs, thereby weakening the cytotox...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/395C07K16/18C07K16/06A61P35/00
Inventor 金坚马鑫陈蕴蔡燕飞何冬旭
Owner JIANGNAN UNIV
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