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Application of EMC8 gene inhibitor on preparation of medicine for treating gastric cancer

A technology for gene suppression and gastric cancer, applied in the field of biomedicine, to achieve the effect of promoting gastric cancer cell apoptosis, inhibiting clone formation, and inhibiting proliferation rate

Active Publication Date: 2019-06-14
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the role of EMC8 gene (ER membrane protein complex subunit 8) in gastric cancer and other malignant tumors has not been reported

Method used

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  • Application of EMC8 gene inhibitor on preparation of medicine for treating gastric cancer
  • Application of EMC8 gene inhibitor on preparation of medicine for treating gastric cancer
  • Application of EMC8 gene inhibitor on preparation of medicine for treating gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Effect of EMC8 gene on the proliferation of gastric cancer cells

[0066] 1.1 Target cells

[0067] The target cells selected in Example 1 are AGS cells (belonging to human gastric adenocarcinoma cells) as shown in Table 3, and the culture medium is a complete medium, that is, F12 medium supplemented with 10% fetal bovine serum.

[0068] Table 3 Example 1 Target cells and culture medium thereof

[0069] target cell

complete medium

96-well plate cell count

AGS cells (gastric adenocarcinoma cells)

F12+10%FBS

1500~2000 pcs / hole

[0070] 1.2 Target virus

[0071] In order to ensure the efficiency of gene interference, in Example 1, 3 RNA interference target sequences were designed for the EMC8 gene. The 3 RNA interference target sequences are shown in Table 4, and 3 plasmid vectors carrying different target sequences were mixed in equal proportions for lentiviral packaging , so as to ensure the knockdown efficiency of the EM...

Embodiment 2

[0119] Example 2 Screening of EMC8 gene inhibitor shRNA and construction of interference lentivirus

[0120] 2.1 Preparation of EMC8 gene interference lentivirus:

[0121] 2.1.1 EMC8 gene target sequence:

[0122] The target sequence of the EMC8 gene is shown in Table 4.

[0123] 2.1.2 Interfering with lentiviral vectors

[0124] Vector name: GV493 (the vector is provided by Jikai Gene Company)

[0125] Element sequence: hU6-MCS-CBh-gcGFP-IRES-puromycin

[0126] Control number: CON313

[0127] Control insert sequence: TTCTCCGAACGTGTCACGT (SEQ ID NO: 4)

[0128] Carrier map such as Figure 4 shown.

[0129] 2.1.3 Design the shRNA interference sequence according to the EMC8 gene target sequence, and synthesize the oligonucleotide chain

[0130] The designed shRNA interference sequence is shown in Table 8, and the shRNA oligonucleotide chain is synthesized according to the sequence, wherein, EMC8-RNAi(70235-11)-a and EMC8-RNAi(70235-11)-b synthesize oligonucleotides Chai...

Embodiment 3

[0172] Example 3 Effects of EMC8 Gene Inhibitors on Gastric Cancer Cells

[0173] MTT experiments, cell apoptosis detection experiments, cell clone formation experiments, and cell cycle detection experiments were carried out to illustrate the effects of EMC8 gene inhibitors on gastric cancer cells, and at the same time prove the effect of the interfering lentivirus constructed in Example 2 on gastric cancer cells. Specifically, unless otherwise specified, the experiments described in Example 3 were conducted using the design in Table 15.

[0174] Table 15 Basic experimental design

[0175]

[0176] 3.1 MTT assay to detect cell viability

[0177] 3.1.1 Experimental steps

[0178] (1) According to 1.4.1-1.4.2 in Example 1, the culture of target cells and lentivirus infection are carried out, and the lentivirus is a lentivirus (NC group) containing a control gene (SEQ ID NO: 4) and Example 2 Prepare the EMC8 gene interference lentivirus LV-EMC8-RNAi (70237-1) (KD group), co...

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Abstract

The invention relates to application of an EMC8 gene inhibitor on preparation of medicine for treating gastric cancer. According to the invention, the EMC8 gene can be used as an effect target for treating the gastric cancer; and the EMC8 gene inhibitor can inhibit the proliferation rate of gastric cancer cells, change the cycle distribution of the gastric cancer cells, promote the apoptosis of the gastric cancer cells and inhibit the clone formation of the gastric cancer cells. The EMC8 gene inhibitor is applied to the preparation of the medicine for treating the gastric cancer, the activityof the EMC8 gene is inhibited at the gene level, or the transcription or expression of the EMC8 gene is inhibited, the prepared medicine for treating the gastric cancer can be a single-component substance or a multi-component substance, the EMC8 gene inhibitor is a unique effective component or one of the effective components, the form of the medicine is not particularly limited, and the medicinecan be in various material forms such as solid, liquid, gel, semifluid, aerosol and the like.

Description

technical field [0001] The invention relates to the use of an EMC8 gene inhibitor in the preparation of a drug for treating gastric cancer, belonging to the field of biomedical research. Background technique [0002] Cancer is one of the major diseases that seriously endanger human health. Among them, gastric cancer is a common malignant tumor of the digestive system. Its incidence rate accounts for the top five new tumors every year in the world, and its mortality rate also ranks among the top five. Gastric cancer mainly originates from malignant tumors of the gastric mucosal epithelium. It ranks among the top incidence rates among various malignant tumors in my country, and the incidence of gastric cancer has obvious regional differences. Globally, the incidence rate in my country is higher than that in Europe and the United States. In my country, the incidence rate of gastric cancer in the northwest and eastern coastal areas is significantly higher than that in the south....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61K31/7105A61K45/06A61P35/00
Inventor 王大志刘新泳展鹏
Owner SHANDONG UNIV
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