Application of miR-588 and target gene thereof in gastric cancer

A technology of mir-588, 1.mir-588, applied in the field of application of miR-588 and its target genes in gastric cancer, can solve the problems of gastric cancer diagnosis, unclear prognosis assessment, and unclear role of miR-588, and achieve Good sensitivity and specificity effect

Pending Publication Date: 2020-11-20
ZHEJIANG CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relationship between miR-588 and gastric cancer has not been reported, and the role of miR-588 in the occurrence and development of gastric cancer is still unclear, especially the relationship between the abnormal expression of miR-588 and the diagnosis and prognosis of gastric cancer is still unclear.

Method used

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  • Application of miR-588 and target gene thereof in gastric cancer
  • Application of miR-588 and target gene thereof in gastric cancer
  • Application of miR-588 and target gene thereof in gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Statistical analysis of miR-588 as a diagnostic molecular marker for gastric cancer

[0068] 1. Comparison and statistical analysis of serum miR-588 expression in patients with gastric cancer and healthy people

[0069] Taking the above-mentioned serum of gastric cancer patients and healthy people as objects, real-time fluorescent quantitative PCR was carried out according to the following method to detect the expression level of serum miR-588 (U6 gene was used as an internal control) in gastric cancer patients and healthy people, and through the scatter diagram to analyze and compare.

[0070] Real-time fluorescent quantitative PCR method:

[0071] Using total RNA as a template for reverse transcription, microRNA is reverse-transcribed into cDNA using specific primers with a stem-loop structure. The product cDNA of RT is detected by quantitative PCR using the TaqMan probe method, and PCR amplification adopts specific pre-primers and universal post-primers ...

Embodiment 2

[0091] Example 2: Prediction and verification of target genes regulated by miR-588

[0092] In order to explore the mechanism of regulation of gastric cancer cells by miR-588 (sequence shown in SEQ ID NO: 1) and find its downstream key target genes, we used the miRWalk database to find thousands of potential target genes. After analysis, we screened CXCL5 (NCBI ID: 6374), CXCL9 (NCBI ID: 4283) and CXCL10 (NCBI ID: 3627) genes. Further, it was also found that: miR-588 directly binds to CXCL5 at the 1171-1177bp site, such as Figure 2A shown; miR-588 directly binds to CXCL9 at the 344-350bp and 914-920bp sites, such as Figure 2B shown. We speculate that miR-588 reduces the viability of gastric cancer cells by interacting with CXCL5, CXCL9 and CXCL10.

[0093] In order to verify the above prediction for the target gene of miR-588, the following experiments were carried out:

[0094] 1. The expression levels of CXCL5, CXCL9 and CXCL10 in gastric tumor tissues and correspondin...

Embodiment 3

[0136] Example 3: Animal experiments confirm that miR-588 is significantly related to the proliferation of gastric cancer cells in vivo

[0137] Six-week-old nude mice (n=15) were purchased from SLAC Experimental Animal Co., Ltd. (Shanghai, China), and were randomly divided into three groups (n=5 in each group): SGC-7901 cells (1×10 7 cells / mL, control group), SGC-7901 cells infected with lentiviral vector (1×10 7 cells / mL, vector group) and SGC-7901 cells infected with miR-588 lentivirus (1×10 7 cells / mL, miR-588 overexpression group), subcutaneously injected into the back of each nude mouse (0.2 mL). Average tumor volumes were measured three times per week. At the end of the experiment (day 35), mice were sacrificed and tumors were excised for volume measurement.

[0138] Figure 4A The photos of the final tumor tissues of mice in each group are given, Figure 4B The data of tumor volume in mice measured every 7 days are given. Figure 4A and Figure 4B It was shown t...

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Abstract

The invention discloses application of miR-588 and a target gene thereof in gastric cancer. It is found that the expression of miR-588 in serum of gastric cancer patients is remarkably lower than thatof miR-588 in serum of healthy people, the expression of miR-588 target genes CXCL5, CXCL9 or CXCL10 in gastric cancer tissues is remarkably lower than that of the healthy people. In the aspect of distinguishing the gastric cancer patients from the healthy people, MiR-588 has better sensitivity and specificity than traditional gastric cancer tumor markers CEA and CA19-9, and miR-588 can be used as a gastric cancer molecular marker; the overexpression miR-588 obviously inhibits the proliferation of gastric cancer cells in vitro and vivo and the vitro invasion by up-regulating the expression oftarget genes CXCL5, CXCL9 or CXCL10; finally, the high-level expression of the miR-588, the CXCL5, the CXCL9 or the CXCL10 is remarkably related to the longer total survival time of a patient suffering from the gastric cancer.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology and biological detection, and specifically relates to the application of miR-588 and its target gene in gastric cancer. Background technique [0002] Gastric cancer ranks second in the mortality rate of malignant tumor patients in the world, and the number of deaths due to gastric cancer is about 1 million worldwide every year. my country is a country with a large incidence of gastric cancer, accounting for about 50% of the world's incidence, and the 5-year survival rate of gastric cancer patients is only 20-30%. For patients with early gastric cancer, surgical resection is the most effective treatment and prolongs the patient's survival time, while advanced gastric cancer often recurs in lymph nodes, blood and peritoneum; in addition, the level of prognosis for patients is still very low. Therefore, early diagnosis of gastric cancer means early intervention and treatment, which is of great...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61K31/7105A61K48/00A61K38/19A61P35/00
CPCC12Q1/6886G01N33/6863G01N33/57446G01N33/57484A61K45/00A61K31/7105A61K48/0008A61K48/005A61K38/195A61P35/00C12Q2600/158C12Q2600/178C12Q2600/118G01N2333/522
Inventor 许颂霄陈勇毅龚万钢
Owner ZHEJIANG CANCER HOSPITAL
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