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Application of human HHIPL2 mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma and kit

A prognostic evaluation and targeted therapy technology, applied in the field of biomedicine, can solve problems such as restricting esophageal cancer

Active Publication Date: 2022-07-29
THE SECOND HOSPITAL OF SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of effective therapeutic targets, the current clinical treatment methods for esophageal cancer are still surgery, radiotherapy and chemotherapy, which seriously restricts the development of individualized precise treatment and comprehensive treatment strategies for esophageal cancer, so that the 5-year survival rate of patients with esophageal cancer Period only about 18%

Method used

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  • Application of human HHIPL2 mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma and kit
  • Application of human HHIPL2 mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma and kit
  • Application of human HHIPL2 mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma and kit

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Experimental program
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Effect test

Embodiment 1

[0034] The TCGA database was used to analyze the sequencing data of esophageal squamous cell carcinoma (ESCC). figure 1 shown.

[0035] The specific implementation process is as follows:

[0036] Download the ESCC specimen sequencing data in the TCGA database and calculate the log of each sample 2 Count value, including 11 cases of adjacent normal esophagus tissue and 95 cases of ESCC tumor tissue samples, with the log of adjacent normal esophagus tissue 2 The average Count value was used as the reference, and the relative expression levels of HHIPL2 mRNA in 95 ESCC tumor tissues were calculated.

Embodiment 2

[0038] siRNA-HHIPL2 was designed, siRNA-HHIPL2 and siRNA-NC were transfected into KYSE-150 cells, respectively, and the knockdown level of HHIPL2 mRNA in KYSE-150 cells was detected by reverse transcription-real-time fluorescence quantitative PCR. The siRNA-HHIPL2 The target sequence is shown in SEQ ID NO.2, the substance used to detect the expression level of HHIPL2 mRNA is the primer pair provided by the present invention, the upstream primer sequence is shown in SEQ ID NO.3, and the downstream primer sequence is SEQ ID NO.4 shown, the specific results are as follows figure 2 shown.

[0039] Depend on figure 2 It can be seen that siRNA-HHIPL2 can effectively and specifically knock down the expression level of HHIPL2 mRNA in KYSE-150 cells.

[0040] The specific implementation process is as follows:

[0041] (1) Design an siRNA (siRNA-HHIPL2) that can target the HHIPL2 mRNA coding region sequence, and its targeting sequence is 5'-cacaatcgcaagttctata-3' (SEQ ID NO.2);

...

Embodiment 3

[0052] KYSE-150 cells were cultured in 6-well plates, and siRNA-HHIPL2 (SEQ ID NO. 2) that specifically interferes with HHIPL2 mRNA expression was transfected into KYSE-150 cells, and the knockdown expression of HHIPL2 mRNA was detected by CCK-8 assay The effect on the proliferation ability of KYSE-150 cells, the specific results are as follows image 3 shown.

[0053] Depend on image 3 It can be seen that the specific knockdown of HHIPL2 mRNA expression level by siRNA-HHIPL2 can significantly inhibit the proliferation of esophageal squamous cell carcinoma cells.

[0054] The specific implementation process is as follows:

[0055] (1) siRNA-HHIPL2 and siRNA-NC were respectively transfected into KYSE-150 cells. The specific method was shown in Example 2. 24-36 hours after transfection, the cells were collected, and the cell concentration was adjusted to 25,000 cells / mL medium. Add 100 μL of cell suspension to each well of the 96-well plate, set 6 duplicate wells for each ce...

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Abstract

The invention relates to application of human HHIPL2mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma and a kit. According to the invention, abnormal high expression of HHIPL2mRNA in esophageal squamous cell carcinoma tumor tissues is found for the first time, and malignant biological behaviors of esophageal squamous cell carcinoma cell line KYSE-150 cells can be remarkably inhibited by knocking down the expression level of HHIPL2mRNA. The kit comprises siRNA capable of specifically interfering with HHIPL2mRNA expression and a primer pair for detecting the HHIPL2mRNA expression level, and the application of the siRNA and the primer pair in esophageal squamous cell carcinoma targeted therapy strategy development is provided.

Description

technical field [0001] The invention relates to an application and a kit of human HHIPL2 mRNA in targeted therapy and prognosis evaluation of esophageal squamous cell carcinoma, and belongs to the technical field of biomedicine. Background technique [0002] Esophageal cancer is a highly aggressive human malignant tumor, which can be divided into esophageal adenocarcinoma and esophageal squamous cell carcinoma (ESCC) according to pathological types. The incidence rates of the two pathological types of esophageal cancer vary greatly in different regions. In western countries, esophageal adenocarcinoma is the main type, while in East Asia, especially my country, ESCC is the most common confirmed case of esophageal cancer. Due to the lack of effective therapeutic targets, the current clinical treatment methods for esophageal cancer are still surgery, radiotherapy and chemotherapy. period is only about 18%. Therefore, there is an urgent need to develop new effective targets fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11C12N15/113A61P35/00A61K31/713
CPCC12Q1/6886C12Q1/6851C12N15/1135A61K31/713A61P35/00C12N2310/14C12Q2600/178C12Q2600/118C12Q2600/158C12Q2525/161C12Q2561/113C12Q2563/107C12Q2531/113
Inventor 赵小刚李培超田忠献郭嘉仲赵云鹏李佩蔚杨凌霄刘繁荣
Owner THE SECOND HOSPITAL OF SHANDONG UNIV
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