Tumour chemotherapy drug sensitive gene and application of same

A chemotherapeutic drug and gene technology, applied in the direction of antineoplastic drugs, plant gene improvement, application, etc., can solve the problem of TJP1 expression loss and other problems, achieve the effect of reducing toxic and side effects, increasing the cure rate, and reducing the number of times of chemotherapy

Inactive Publication Date: 2011-10-19
PERSONGEN BIOMEDICINESUZHOUCO
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  • Claims
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Problems solved by technology

In the Velcade drug-resistant cell line RPMI 8226/BR we established, quant

Method used

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  • Tumour chemotherapy drug sensitive gene and application of same
  • Tumour chemotherapy drug sensitive gene and application of same
  • Tumour chemotherapy drug sensitive gene and application of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Correlation analysis between the expression of TJP1 gene and the effect of chemotherapy in tumor patients.

[0033]Through the high-throughput and large-scale RNA interference library screening work we established, we screened TJP1 as a potential chemotherapeutic drug-sensitive gene. If TJP1 determines the sensitivity of tumor patients to chemotherapy drugs, then patients with positive expression of TJP1 should respond to chemotherapy drugs, otherwise they will be insensitive to chemotherapy drugs. To this end, we performed gene map expression analysis to determine whether TJP1 gene expression was significantly correlated with patient outcomes. The gene expression database we use comes from the Multiple Myeloma Genomics Portal in the United States (http: / / www.broad.mit.edu / mmgp / pages / publicPortalHome.jsf), in which the Millennium database contains 264 cases of myeloma patients who passed Velcade or local Gene expression microarray data after dexamethasone treatment, an...

Embodiment 2

[0035] To quantitatively determine the expression of TJP1 gene in tumor cells.

[0036] Quantitative PCR was used to determine the expression level of TJP1 in tumor cells. Total RNA was prepared from tumor cells using the QIAGEN RNeasy kit (catalogue number 74104), and the preparation steps were strictly followed the product instructions. Wherein, the homogenization of tissues and cells is carried out using QIAshredder of QIAGEN (catalogue number 79654).

[0037] The concentration and quality of the prepared RNA were determined by spectrophotometry, and 1 μg of RNA was used for eDNA synthesis. The cDNA synthesis kit was carried out using Invitrogen's SuperScript III First-Strand Synthesis Super Mix (catalogue number 18080-400), and the synthesis steps were strictly followed the product instructions.

[0038] When performing quantitative PCR reaction, dilute the synthesized cDNA 5 times with nuclease-free deionized water, take 1 μl of diluted cDNA in 0.2ml In the Optical re...

Embodiment 3

[0041] The expression of TJP1 gene was significantly correlated with the semi-lethal dose (IC50) of chemotherapeutic drugs in tumor cells.

[0042] We compared the IC50 of representative MOLP-8 (no TJP1 expression) and MM1.s (high TJP1 expression) cell lines. The specific steps of the experiment were: WST-1 (purchased from Roche Diagnostics) was used to determine the IC50 data were obtained from the effects of mycin and cisplatin on cell proliferation. Cells were seeded in a 96-well culture plate at a density of 2x104 / well, and were exposed to different concentrations of doxorubicin and cisplatin after 24 hours, and the control cells were treated with a drug-free solvent. After 48 hours, the cell proliferation was measured with WST-1, and the measurement method was strictly implemented according to the product instructions. It was found that the IC50 of MM1.s cells expressing TJP1 to doxorubicin and cisplatin (38.2nM and 13nM respectively) were significantly lower than those ...

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Abstract

The invention discloses a gene capable of determining the sensitivity of tumour cells to chemotherapy drugs, which is named TJP1 or ZO-1. The invention also discloses a method for predicting the chemotherapeutic effect on tumour by using the gene, and patients in which TJP1exrpression exists are sensitive to the chemotherapy drugs; and the patients suffering the loss of expression of TJP1 are endowed with tolerance to the chemotherapy drugs. The invention also discloses a method for improving the chemotherapeutic effect by improving or inducing TJP1 expression as well as the application of the gene to the prediction of chemotherapeutic effect on tumour, the increasing of tumour chemotherapy sensitivity and the preparation of oncotherapy drugs. The invention can be beneficial to the development of a TJP1/ZO-1 expression related diagnostic reagent kit so as to predict the chemotherapeutic effect on patients, and also beneficial to the screening of anticarcinogen so as to find out chemical and biological molecules inducing the TJP1/ZO-1 expression, thereby finally achieving the purposes of reducing the times of chemotherapy, increasing the recovery rate, alleviating the toxic side effects of the chemotherapy drugs, saving the hospitalization costs and generating outstanding economic and social benefits.

Description

technical field [0001] The present invention relates to a gene that plays a decisive role in the sensitivity of tumor cells to chemotherapeutic drugs and its application, in particular to detecting the gene as a target to predict whether a patient is sensitive to chemotherapeutic drugs, and to induce or increase the gene's sensitivity in the patient's body. Expression in order to enhance the effect of chemotherapy in patients. Background technique [0002] The main treatment methods for cancer are surgical resection, radiation therapy, chemical drug therapy and immunological therapy. Among them, chemotherapy-chemotherapy is a systemic treatment method for tumors. It has been widely used and has become the main method for the treatment of malignant tumors, especially for hematological tumors, such as leukemia, lymphoma and myeloma, as well as the treatment of middle and advanced cancers. However, tumor chemotherapy drugs have great toxic side effects and are prone to drug re...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12Q1/68A61K48/00A61P35/00
Inventor 杨林杨楠黄行许吴小军钱建飞杨文荣
Owner PERSONGEN BIOMEDICINESUZHOUCO
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