Application of IL-11 monoclonal antibody in inhabiting PI3K/Akt signal channel

A monoclonal antibody, 1. IL-11 technology, applied in the direction of antibodies, antineoplastic drugs, peptides, etc., can solve side effects and other problems, and achieve the effect of reducing toxic and side effects

Active Publication Date: 2016-04-20
HEILONGJIANG HEIKE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide an application of IL-11 monoclonal antibody in inhibiting PI3K

Method used

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  • Application of IL-11 monoclonal antibody in inhabiting PI3K/Akt signal channel
  • Application of IL-11 monoclonal antibody in inhabiting PI3K/Akt signal channel
  • Application of IL-11 monoclonal antibody in inhabiting PI3K/Akt signal channel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1I

[0017] Example 1 The effect of IL-11 on the proliferation of intestinal cancer cells

[0018] 1. Screening of IL-11 concentration

[0019] Dilute the digested HT-29 cell suspension of the 3rd to 5th generation by 10 4 / ml concentration inoculated in 12-well culture plate, 2ml per well, placed at 37°C, 5% CO 2 cultured in an incubator. After 12 hours of adherence, the culture medium was discarded, and 2 ml of DMEM complete medium with a final concentration of (2, 5, 8, 10 ug / ml) IL-11 was added, and a cell control without IL-11 and a repeated control were established at the same time. Continue culturing, add 80ul MTT to the cells at 6h, 12h, 24h, 36h, and 48h, add 150ul dimethyl sulfoxide after continuing to culture for 4h, shake for 10 minutes, and detect OD490.

[0020] 2. The effect of IL-11 on cell proliferation

[0021] cell suspension according to l0 4 / ml concentration inoculated in 12-well culture plate, 2ml per well, placed at 37°C, 5% CO 2 cultured in an incubat...

Embodiment 2I

[0026] The effect of embodiment 2IL-11 on PTEN, AKT, NF-κB in PI3K cell signaling pathway

[0027] Cell suspension by 5xl0 4 Inoculate in 12-well culture plate (with slide), 2ml per hole, at 37°C, 5% CO 2 cultured in an incubator. After 12 hours of attachment, the culture medium was discarded, and the experiment was divided into two groups. One group was added with 2ml of DMEM complete medium with a final concentration of (8ug / ml) IL-11. After 4h, a volume of 10ulIL-11 antibody was added to incubate for 4h. One group only added 2ml of DMEM complete medium with a final concentration of (8ug / ml) IL-11, and a signal inhibitor control group and a repeat group were set up at the same time. Continue to culture, collect cell supernatant from 4h, 6h, 8h, 12h, detect PTEN, AKT with ELISA kit;

Embodiment 3

[0028] Embodiment 3 detects the content of PTEN in the above-mentioned cell supernatant

[0029] 1. Method:

[0030] 1) Using an ELISA kit to detect the content of PTEN in the supernatant of the above cells.

[0031] 1. Adding samples: set blank wells, standard wells, and sample wells to be tested respectively. Add 100ul of sample diluent to the blank well, and add 100ul of standard substance or sample to be tested in the remaining wells respectively. Be careful not to have air bubbles. Cover or cover the target plate and react at 37°C for 120 minutes.

[0032] 2. Discard the liquid, spin dry without washing. Add 100ul of detection solution A working solution to each well (prepared by taking 1ul detection solution A plus 99ul detection diluent A, mix gently, and prepare within one hour before use), 37°C, 60 minutes.

[0033] 3. After incubating for 60 minutes, discard the liquid in the well, spin dry, wash the plate 3 times, soak for 1-2 minutes each time, 350ul / well, and ...

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Abstract

The invention discloses the application of an IL-11 monoclonal antibody in inhabiting a PI3K/Akt signal channel. The concentration of the IL-11 monoclonal antibody is 8 mug/mL, and the dosage of the IL-11 monoclonal antibody is 10 mug. The safe and efficient PI3K/Akt signal channel inhibitor, namely the IL-11 monoclonal antibody is found, and can be specifically combined with IL-11 secreted by cells to induct PTEN transcription, up-regulate PTEN expression and inhibit the expression of AKT on the downstream portion of the PI3K/Akt signal channel, and then the whole signal channel is inhibited. The inhibitor is a natural antibody, so that toxic and side effects are reduced and safety and efficiency are high.

Description

technical field [0001] The invention belongs to the technical field of intracellular signaling pathways, and specifically relates to the application of an IL-11 monoclonal antibody in inhibiting PI3K / Akt signaling pathways. Background technique [0002] Colon cancer is one of the common malignant tumors with a five-year survival rate of less than 5%. Therefore, research on the molecular mechanism of colon cancer development has attracted much attention. The occurrence of colon cancer is a multi-factor, multi-stage and multi-gene mutation process, involving molecular events mediated by multiple proto-oncogenes and tumor suppressor genes. In recent years, with the development of cell biology and molecular biology, people have recognized The expression products of most oncogenes are components in the cell signal transduction pathway, and they can interfere with the cell signal transduction process from multiple links, leading to the proliferation and differentiation of tumor c...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P35/00
CPCA61K2039/505C07K16/244
Inventor 曹涤非黄国庆吴琼赵金海薛佳莹王雷王东凯孙尧李瑶
Owner HEILONGJIANG HEIKE TECH CO LTD
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