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Use of polyphosphoinositide phosphatase 1 as a target molecule

An overexpression and gene technology, applied in the field of polyphosphoinositide phosphatase 1 as a target molecule, can solve the problems that there are no related reports and no related research on the regulatory mechanism, and achieve the goal of inhibiting growth and invasion and inhibiting proliferation Effect

Active Publication Date: 2022-02-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the expression of MINPP1 in patients with HBV-related hepatocellular carcinoma and the regulatory mechanism of MINPP1 after HBV infects host cells have not been reported yet.
In addition, whether MINPP1 can be used as a new target for anti-HBV-related hepatocellular carcinoma has not yet been studied.

Method used

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  • Use of polyphosphoinositide phosphatase 1 as a target molecule
  • Use of polyphosphoinositide phosphatase 1 as a target molecule
  • Use of polyphosphoinositide phosphatase 1 as a target molecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Determination of the expression of MINPP1 in HBV-related and non-HBV-related hepatocellular carcinoma

[0038] Tumor tissues from 24 patients with HBV-related HCC and 24 patients with non-HBV-related HCC were collected through surgical resection, and the specimens were collected and placed in a -80°C refrigerator for later use. When extracting the RNA in the tissue, take the sample out of the low-temperature refrigerator and thaw it, add 1mL Trizol reagent per 100mg, homogenize with an electric homogenizer, and let it stand for 5min after the end. Add 200 μL of chloroform to the sample, shake vigorously, let it stand for 5 minutes, and centrifuge at 12,000 rpm for 15 minutes in a centrifuge precooled to 4°C. After centrifugation, the sample can be separated. Transfer the upper aqueous phase to a new centrifuge tube, add an equal volume of isopropanol and mix gently, and let it stand at room temperature for 20-30min. Centrifuge the sample in the centrifuge tu...

Embodiment 2

[0043] Example 2: MINPP1 gene can inhibit the growth and invasion of HBV-related liver cancer cells

[0044] In this example, one HBV-related liver cancer cell line, Hep3B, and one non-HBV-related liver cancer cell line, Huh7, were respectively selected and plated on cells containing DMEM medium (containing 10% fetal bovine serum and 1% penicillin / streptomycin) Petri dish and placed in a carbon dioxide incubator (37°C, 5% CO 2 ) cultured until the number of cells is about 1×10 6 cells / mL were collected. Adopt the same method as in Example 1 to extract the RNA of the cells, and carry out RT-PCR and qPCR experiments to measure the relative expression of MINPP1 in two strains of cells, the results are as follows Figure 4 As shown in A, it can be seen that the expression level of MINPP1 gene in Hep3B cells was significantly lower than that in Huh7 cells under the same culture conditions.

[0045] Further, the effect of MINPP1 on the growth of HBV-related liver cancer cells was...

Embodiment 3

[0048] Example 3: MINPP1 can inhibit tumor growth in animal models

[0049] In this example, firstly, the plasmid capable of overexpressing MINPP1 was transfected into Hep3B cells and Hep3B cells were used as a control. After transfection, the cells were collected and resuspended in serum-free DMEM medium, adjusted to a cell concentration of 2×10 7 cell / mL, inoculate 100 μL of cell suspension into nude mice subcutaneously with a syringe, three mice in each group. After inoculation, the tumor formation in nude mice was observed and the tumor volume was recorded. On the 30th day after inoculation, the nude mice with tumor formation were sacrificed by neck dislocation, and the tumors were taken off and recorded. The result is as Figure 6 As shown in A, it can be seen that the tumor volume on the side of inoculation on the back of nude mice gradually increases with time. The growth rate of the tumor in the nude mice in the control group was significantly faster on the 13th day ...

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Abstract

The invention discloses the application of MINPP1 as a target molecule in the preparation of anti-HBV-related hepatocellular carcinoma drugs, and inhibits the growth and invasion of HBV-related hepatocellular carcinoma cells by overexpressing or activating the MINPP1 gene. The present invention confirms that the MINPP1 gene can be used as an important target for effectively inhibiting the proliferation of HBV-related liver cancer cells.

Description

technical field [0001] The invention relates to the application of a multiple inositol polyphosphate phosphatase 1 (Multiple Inositol PolyphosphatePhosphatase 1, MINPP1) gene as a target for anti-hepatitis B virus-related hepatocellular carcinoma. Background technique [0002] Primary liver cancer is caused by the canceration of liver-related cells in the body. It has the characteristics of high malignancy, easy recurrence and metastasis, and poor prognosis, which seriously endangers human health. According to statistics, in 2018, there were about 841,000 new cases of liver cancer worldwide, and about 781,000 deaths. my country is a country with a high incidence of liver cancer, with approximately 460,000 new cases each year. According to histopathological classification, primary liver cancer can be divided into hepatocellular carcinoma, intrahepatic cholangiocarcinoma and mixed cell carcinoma. Among them, hepatocellular carcinoma (HCC) is a malignant tumor of liver epithe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61P35/00A61P1/16
CPCA61K45/00A61P35/00A61P1/16
Inventor 刁宏燕王凯航陈文标姜晶晶章旭君毕珂凡
Owner ZHEJIANG UNIV
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