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Application of gbp1 gene and its inhibitor in preparation of medicine for treating lung squamous cell carcinoma

A technology for gene suppression and squamous cell carcinoma of the lung, applied in gene therapy, drug combination, antineoplastic drugs, etc., can solve problems that have not been confirmed by research, increased gene expression, etc.

Active Publication Date: 2018-11-16
江苏天芳健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The applicant's previous research found that compared with normal lung tissue, the expression of GBP1 gene in lung squamous cell carcinoma pathological tissue was significantly increased, but so far no research has confirmed the relationship between GBP1 and lung squamous cell carcinoma

Method used

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  • Application of gbp1 gene and its inhibitor in preparation of medicine for treating lung squamous cell carcinoma
  • Application of gbp1 gene and its inhibitor in preparation of medicine for treating lung squamous cell carcinoma
  • Application of gbp1 gene and its inhibitor in preparation of medicine for treating lung squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 miR-335-3p targeted down-regulation of GBP1 gene expression

[0022] 1. Experimental materials

[0023] miR-335-3p mimics miR-335-3p mimics, repressor miR-335-3p inhibitor, miR-335-3p mimics negative control miR-335-3p mimics negative control and repressor negative control miR-335-3pinhibitor Negative control was purchased from Guangzhou Ruibo Biotechnology Co., Ltd.

[0024] The PCR primers were designed and synthesized by Guangzhou Ruibo Biotechnology Co., Ltd.

[0025] TRIzol reagent and transfection reagent Lipofectamine 2000 were purchased from Invitrogen.

[0026] The NCI-H520 cell line of lung squamous cell carcinoma was stored in our company for a long time, and the Hela cell line used for transfection of the plasmid vector was purchased from the Cell Bank of Shanghai Institute of Cell Biology, Chinese Academy of Sciences. RPMI1640 medium and fetal bovine serum were purchased from Gibco. Lung squamous cell carcinoma NCI-H520 cell line and Hela cell...

Embodiment 2

[0072] Example 2 Discovery of small molecule compounds targeting down-regulation of GBP1 gene expression

[0073] 1. Experimental materials

[0074] Siberimine (CAS No.: 61825-98-7), Sylphine (CAS No.: 518-94-5) and Buicilline (CAS No.: 860-79-7) are stored in the standard library of our company compound of. Dissolve siberimine, cylindrine and epiphylline in dimethyl sulfoxide to make a mother solution with a concentration of 1 mg / mL, and dilute it with the medium to the required concentration when used.

[0075] Other experimental materials are the same as in Example 1.

[0076] 2. Experimental method

[0077] 1. Cell culture and compound administration

[0078] Lung squamous cell carcinoma NCI-H520 cell line was cultured in RPMI1640 medium containing 10% fetal bovine serum according to conventional methods. The NCI-H520 cells were divided into 7 groups, namely the low-dose group and high-dose group of siberimine, the low-dose group and high-dose group of sylvaline, the ...

Embodiment 3

[0105] Example 3 In vivo tumor-inhibitory effect of small molecule compounds targeting down-regulation of GBP1 gene expression on lung squamous cell carcinoma

[0106] 1. Experimental materials

[0107] SPF grade BALB / c nude mice were purchased from Shanghai Slack Experimental Animal Co., Ltd., 7 weeks old, weighing 19.5-22.5 g. All mice were reared in an environment with temperature (22±2)°C, humidity (60±5)°C, and alternating light and dark for 12 hours.

[0108] 2. Experimental method

[0109] 1. Establishment, grouping and administration of animal models

[0110] The lung squamous cell carcinoma NCI-H520 cells in the logarithmic growth phase were digested with 0.25% trypsin to make a single cell suspension. The number of living cells accounted for more than 95% by the trypan blue exclusion method, and the cell density was adjusted to 1×10 7 / ml, 0.2ml of the cell suspension was inoculated subcutaneously on the dorsal side of the right thigh of each nude mouse, and the b...

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Abstract

The invention discloses a GBP1 gene and applications of a GBP1 gene inhibitor in preparation of a drug for treating lung squamous carcinoma. The expression level of GBP1 gene in lung squamous carcinoma NCI-H520 cells can be obviously down-regulated by miR-335-3p, dual-luciferase report gene detection results show that GBP1 gene can be directly regulated by miR-335-3p. Cell proliferation activity detection results show that in-vitro proliferation of the lung squamous carcinoma NCI-H520 cells can be remarkably inhibited by miR-335-3p through down-regulating the expression of the GBP1 gene, and the miR-335-3p is an inhibitor for expression of the GBP1 gene. Sipeimine, cycleanine and cyclovirobuxine have an obvious inhibition effect on the growth of lung squamous carcinoma NCI-H520 cell transplantation tumor, the expression level of GBP1 mRNA and protein in tumor tissues treated by sipeimine, cycleanine and cyclovirobuxine can be remarkably down-regulated, thus showing that the expression of the GBP1 gene can promote the growth and proliferation of the lung squamous carcinoma, and the in-vivo tumor inhibition effect of the sipeimine, cycleanine and cyclovirobuxine can be realized by down-regulating the expression of the GBP1 gene.

Description

technical field [0001] The invention belongs to the field of gene therapy drugs, and in particular relates to the application of GBP1 gene and its inhibitors in the preparation of drugs for treating lung squamous cell carcinoma. Background technique [0002] Lung cancer is the most common malignant tumor in the world, and its mortality rate ranks first among malignant tumors. The incidence of lung cancer in my country has been increasing year by year, with an average annual increase of nearly 2%. Lung cancer is divided into multiple tissue types according to different pathological characteristics, and the treatment measures are also different for different types of lung cancer tissue. According to the 2004 edition of the WHO classification, common histopathological types of lung cancer are divided into non-small cell carcinoma (NSCLC) and small cell carcinoma (SCLC). Non-small cell carcinomas are subdivided into squamous cell carcinoma (SCC), adenocarcinoma (AC) and large ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K45/00A61P35/00A61K31/58A61K31/4748A61K31/57
CPCA61K31/4748A61K31/57A61K31/58A61K45/00A61K48/005
Inventor 郭守河张红
Owner 江苏天芳健康科技有限公司
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