SiRNA, siRNA plasmid and lentivirus for targeted inhibition of neutral cholesteryl ester hydrolase 1 (NCEH1) gene expression, and construction methods and application of SiRNA, siRNA plasmid and lentivirus

A technology of gene expression and lentivirus, which is applied in the field of genetic engineering, can solve the problems of short-lived gene inhibition and short half-life of siRNA fragments, and achieve the effects of inhibiting proliferation, promoting apoptosis of liver cancer cells, inhibiting growth, proliferation, and invasion and metastasis

Active Publication Date: 2021-02-23
广州市红十字会医院
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Problems solved by technology

However, due to the short half-life of siRNA fragments, after the siRNA fragments synthesized i

Method used

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  • SiRNA, siRNA plasmid and lentivirus for targeted inhibition of neutral cholesteryl ester hydrolase 1 (NCEH1) gene expression, and construction methods and application of SiRNA, siRNA plasmid and lentivirus
  • SiRNA, siRNA plasmid and lentivirus for targeted inhibition of neutral cholesteryl ester hydrolase 1 (NCEH1) gene expression, and construction methods and application of SiRNA, siRNA plasmid and lentivirus
  • SiRNA, siRNA plasmid and lentivirus for targeted inhibition of neutral cholesteryl ester hydrolase 1 (NCEH1) gene expression, and construction methods and application of SiRNA, siRNA plasmid and lentivirus

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[0036] Figure 1 to Figure 10 Shown are examples of siRNA, siRNA plasmid, interfering lentivirus and its construction method and application for targeting and inhibiting the expression of NCEH1 gene of the present invention, by studying the expression level of NCEH1 in human liver cancer cell lines, and designing the siRNA sequence of human NCEH1 gene , and construct the corresponding siRNA plasmid and interfering lentivirus, and use the lentivirus-mediated siRNA method to specifically inhibit the expression of NCEH1 gene in human liver cancer cells, thereby inhibiting the proliferation, invasion and metastasis of human liver cancer cells, and play a role in the treatment of liver cancer.

[0037] 1. Real-time quantitative PCR to detect the expression level of NCEH1 gene in cell lines

[0038] 1) Cell culture: HL7702 normal human liver cell line and SK-HEP-1, Bel-7402, SMMC-7721, HepG2 and Hep3B human liver cancer cell lines were placed at 37°C, 5% CO 2 The cells were culture...

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Abstract

The invention discloses siRNA, siRNA plasmid and lentivirus for targeted inhibition of expression of neutral cholesteryl ester hydrolase 1 (NCEH1) gene of a human hepatocellular carcinoma cell, construction methods of the siRNA, the siRNA plasmid and the lentivirus, and application of the siRNA, the siRNA plasmid and the lentivirus to preparation of hepatocellular carcinoma treatment drugs for inhibiting proliferation, invasion and metastasis of the hepatocellular carcinoma cell. By researching the expression level of the NCEH1 in hepatocellular carcinoma tissue, such action target, the NCEH1gene, is provided for treatment of hepatocellular carcinoma; by designing the sequence of the siRNA for the targeted inhibition of the expression of the NCEH1 gene of the human hepatocellular carcinoma cell, the corresponding siRNA expression plasmid and siRNA lentivirus, the expression of the NCEH1 gene in the human hepatocellular carcinoma cell can be efficiently inhibited, and thus the proliferation, invasion and metastasis capability of the hepatocellular carcinoma cell is effectively inhibited; the siRNA, siRNA plasmid and lentivirus are of great significance in preparation of drugs for inhibiting proliferation, invasion and metastasis of the hepatocellular carcinoma cell so as to treat the hepatocellular carcinoma, and an effective method is provided for developing targeted therapy drugs for treating the hepatocellular carcinoma for NCEH1.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to siRNA, siRNA plasmid, lentivirus and its construction method for targeting and inhibiting the expression of NCEH1 gene in human liver cancer cells, and in the preparation of liver cancer therapeutic drugs for inhibiting the proliferation, invasion and metastasis of liver cancer cells Applications. Background technique [0002] Hepatocellular Carcinoma (HCC; liver cancer for short) now ranks fourth in the incidence of malignant tumors in my country, and its annual mortality rate has reached 20.37 / 100,000, ranking third in the mortality rate of malignant tumors in my country, seriously threatening the people of our country. life and health. Although the surgical resection rate of liver cancer has increased significantly in recent years, the treatment methods for liver cancer have become more diversified, and liver transplantation is increasingly used in the trea...

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Application Information

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IPC IPC(8): C12N15/113C12N15/867C12N15/66A61K48/00A61K31/713A61P35/00
CPCC12N15/1137C12N15/86C12N15/66A61K31/713A61P35/00C12N2310/141C12N2740/15043C12N2800/107C12Y301/01079
Inventor 吴帆王百林曾裕文
Owner 广州市红十字会医院
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