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New application of aldehyde dehydrogenase 2 gene

An acetaldehyde dehydrogenase and application technology, applied in the field of medical detection, can solve the problems of less chemotherapy drugs, lag in targeted therapy, and high heterogeneity of gastric cancer

Active Publication Date: 2022-06-21
天筛(上海)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Judging from the current situation, there are few chemotherapy drugs to choose from (only three major categories), targeted therapy lags behind (trastuzumab is only suitable for about 15% of patients), and gastric cancer has high heterogeneity. These three items are advanced gastric cancer The main treatment bottleneck of

Method used

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  • New application of aldehyde dehydrogenase 2 gene
  • New application of aldehyde dehydrogenase 2 gene
  • New application of aldehyde dehydrogenase 2 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Establishment of gastric cancer cell line with ALDH2 knockdown expression

[0048] Human gastric cancer cell SGC-7901 was purchased from the cell bank of Shanghai Institute of Cell Biology, Chinese Academy of Sciences, and embryonic kidney cell 293T was deposited by the institute. Cell culture conditions: SGC-7901 was cultured with RPMI-1640 cell culture medium containing 10% calf serum at 37°C and 5% CO2, and cells in logarithmic growth phase were used for experiments.

[0049] The retrovirus vector for ALDH2 knockdown expression was constructed, and the steps of the retrovirus packaging method were the same as described above. SGC-7901 cells were infected with retrovirus for 48 hours, and the final concentration of 1 μg / ml puromycin was used for screening to obtain a stable knockdown expression cell line , the knockdown effect was verified by qRT-PCR, Western Blot and immunofluorescence, respectively. The results showed that small interfering RNA could sig...

Embodiment 2

[0051] Example 2: Drug sensitivity test of ALDH2 knockdown gastric cancer cells

[0052] The sensitivity of gastric cancer cells to 5-Fu, cisplatin and DOX by knocking down the ALDH2 gene was compared. We treated the ALDH2 knockdown expressing gastric cancer cells SGC-7901 (ALDH2 shRNA1 and shRNA2 cells) and the control group from Example 1 with several chemotherapeutic drugs at gradient concentrations (0 μM, 0.01 μM, 0.1 μM, 1 μM, 10 μM and 100 μM concentrations), respectively. Gastric cancer cell SGC-7901 (shRNA empty vector), the relative survival rate of cells in each group was compared.

[0053] The study showed that the relative survival rate of ALDH2 shRNA1 and shRNA2 cells was significantly decreased in gastric cancer cells treated with DOX for 24 h compared with the shRNA empty vector group (P=0.0014, P=0.0082), suggesting that ALDH2 knockdown cells were more sensitive to DOX treatment. Gastric cancer cells treated with cisplatin for 24h, ALDH2-shRNA1 and ALDH2-shRNA...

Embodiment 3

[0059] Example 3: Study on drug sensitivity of tumor xenografts inoculated with ALDH2 gene knockdown gastric cancer cells in nude mice

[0060] To evaluate the difference in drug sensitivity of ALDH2 gene-deficient gastric cancer cells in vivo, we performed tumor inoculation and experimental treatment studies using immunodeficient nude mice. We used a total of 14 4-week-old BALB / C nude mice, inoculated subcutaneously with 3×106ALDH2-shRNA1 and shRNA empty vector SGC-7901 cells in the left and right thighs of the nude mice, respectively, and the transplanted tumors grew to 150mm. 3 , 7 rats in the experimental group and 7 in the control group were given intraperitoneal injection of 2 mg / Kg body weight of DOX and DMSO, twice a week, the tumor size was measured before each drug injection, treated for 3 consecutive weeks, and 3 days after the last drug injection The tumor size was measured and the mice were sacrificed. The study found that the tumor weight of the subcutaneously tr...

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Abstract

The invention belongs to the field of medical detection, and in particular relates to a new application of acetaldehyde dehydrogenase 2 gene. The invention discloses the use of the acetaldehyde dehydrogenase 2 gene as a detection index for gastric cancer molecular typing, and also discloses the use of anthracyclines in the preparation of acetaldehyde dehydrogenase 2 gene mutation or deletion type gastric cancer treatment drugs. For the first time, it was found that the deletion or mutation of the ALDH2 gene caused gastric cancer cells to show different sensitivities to tumor chemotherapy drugs, and the in vivo experiments were verified, providing a new detection index for the molecular typing of gastric cancer. The application of the aldehyde dehydrogenase 2 gene mutation or deletion type gastric cancer treatment drug provides a new option for individualized treatment of gastric cancer.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a new application of acetaldehyde dehydrogenase 2 gene. Background technique [0002] Gastric cancer is the fourth most common cancer worldwide and the second leading cause of death. The cases in East Asia account for more than half of the total, and the incidence in China accounts for 43% of the global incidence and the mortality rate accounts for 45% of the world. Moreover, Chinese gastric cancer patients were found at a late clinical stage, and the proportion of advanced gastric cancer accounted for about 70%, which is the most concerned group of people. Because of the strong heterogeneity of gastric cancer, the prognosis and treatment outcome of gastric cancer are different depending on the location, case type, age, gender, etc. of gastric cancer. [0003] The classification of gastric cancer has undergone gross morphological typing, histological typing, environm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K31/704A61K45/06A61P35/00
CPCA61K45/06A61K31/704C12Q1/6886C12Q2600/136C12Q2600/156
Inventor 宋晓英
Owner 天筛(上海)科技有限公司
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