Long-chain non-coding RNA and application thereof in diagnosis and treatment of hepatocellular carcinoma

A long-chain non-coding, hepatocellular carcinoma technology, applied in the treatment of hepatocellular carcinoma, long-chain non-coding RNA and its application in the field of diagnosis

Inactive Publication Date: 2021-08-17
ZHEJIANG MEDICAL COLLEGE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, as a malignant tumor with extremely high morbidity and mortality, hepatocellular carcinoma currently has certain limitations in terms of clear diagnosis, guiding treatment, and judging prognosis no matter whether it is hematology, imaging, or histology.

Method used

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  • Long-chain non-coding RNA and application thereof in diagnosis and treatment of hepatocellular carcinoma
  • Long-chain non-coding RNA and application thereof in diagnosis and treatment of hepatocellular carcinoma
  • Long-chain non-coding RNA and application thereof in diagnosis and treatment of hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] This example provides verification of the expression and prognosis of lncRNA MCM3AP-AS1 in HCC.

[0046] 1. Experimental materials and methods

[0047] 1.1 Clinical specimens

[0048] A total of 80 pairs of HCC and paracancerous tissues were collected from patients undergoing hepatectomy. Sample collection criteria were as follows: ① confirmed as HCC by two pathologists; ② did not receive any preoperative treatment; ③ not associated with other cancers. All samples were washed with pre-cooled PBS and immediately frozen in liquid nitrogen and stored in a -80°C refrigerator. 80 patients were followed up to collect survival data.

[0049] 1.2 Cell lines and culture

[0050] Immortalized human normal liver cells LO2 and HCC cell lines (Hep3B, HepG2, Huh7, SMMC-7721) were purchased from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). Hep3B was cultured in MEM medium (Hyclone, USA), and LO2, HepG2, Huh7, SMMC-7721 were cultured ...

Embodiment 2

[0069] This example discusses the effect of interfering lncRNA MCM3AP-AS1 on the biological function of HCC cells (Hep3B, HepG2).

[0070] 1. Experimental materials and methods

[0071] 1.1 Cell lines and culture

[0072] Same as Example 1

[0073] 1.2 RNA fluorescence in situ hybridization (Fluorescence in situ hybridization, FISH)

[0074] Cy3-labeled lncRNA MCM3AP-AS1 probe and subcellular localization detection kit were purchased from Ribo (Guangzhou, China), and observed using a confocal laser microscope (Carl Zeiss, Oberkochen, Germany).

[0075] 1.3 Cell Transfection

[0076] Two specific lentivirus-mediated shRNAs (sh-MCM3AP-AS1-1 and sh-MCM3AP-AS1-2) targeting lncRNA MCM3AP-AS1 and the negative control sh-control were designed and synthesized by Jisai Biotech (Guangzhou, China) , and the interference sequences are SEQUENCE NO.4 and SEQUENCE NO.5, respectively. Lentivirus infection of HCC cells was carried out with the transfer promoter Polybrene (8ng / ml).

[007...

Embodiment 3

[0095] 1. Experimental materials and methods

[0096] 1.1 Cell lines and culture

[0097] Same as Example 1

[0098] 1.2 Tumor xenograft model

[0099] 4-5 weeks old, 18-20 g, BALB / C male nude mice (Slack, Shanghai, China) were selected and randomly divided into two groups (n=6 in each group). Hep3B cells transfected with sh-MCM3AP-AS1 and sh-control (1×10 6 ) into the right flank of mice by subcutaneous injection. Tumor volume was measured every three days after tumor formation. Volume calculation formula: volume=(length×width×width) / 2. After 3 weeks, all mice were sacrificed under anesthesia.

[0100] 1.3 Data statistics and analysis

[0101] Same as Example 1

[0102] 2. Experimental results

[0103] 2.1 In Example 3, the present invention established a tumor xenograft model in nude mice by subcutaneously injecting Hep3B cells transfected with sh-MCM3AP-AS1 and sh-control. The results of this in vivo experiment show that knocking down lncRNA MCM3AP-AS1 can inhibit ...

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Abstract

The invention discloses a long-chain non-coding RNA and an application of the long-chain non-coding RNA in diagnosis and treatment of hepatocellular carcinoma. The long-chain non-coding RNA is lncRNA MCM3AP-AS1, and the nucleotide sequence of the lncRNA MCM3AP-AS1 is as shown in Seq ID NO. 1. It is found for the first time that lncRNA MCM3AP-AS1 can be used as a prognosis biomarker and a drug treatment target of hepatocellular carcinoma, lncRNA MCM3AP-AS1 is obviously up-regulated in hepatocellular carcinoma tissues and cell lines, the prognosis of HCC patients overexpressing lncRNA MCM3AP-AS1 is poorer, meanwhile, lncRNA MCM3AP-AS1 has good hepatocellular carcinoma prognosis ability, and through in-vitro and in-vivo experiments, it is proved that the lncRNA MCM3AP-AS1 exists in cytoplasm, and the proliferation of hepatocellular carcinoma can be inhibited by knocking down the lncRNA MCM3AP-AS1, such that the lncRNA MCM3AP-AS1 can provide the cancer promoting effect in the occurrence and development process of hepatocellular carcinoma, and can be adopted as the novel treatment target of hepatocellular carcinoma.

Description

Technical field: [0001] The invention relates to the field of genetic engineering, in particular to a long-chain non-coding RNA and its application in diagnosing and treating hepatocellular carcinoma. Background technique: [0002] According to the 2018 global cancer statistics, for primary liver cancer (PLC), there are approximately 841,000 new cases and 782,000 deaths worldwide. Among them, China accounts for about 50% of the total new cases and total deaths. According to histological types, primary liver cancer includes hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and mixed liver cancer (HCC-ICC). Among them, HCC accounts for about 75-80% of PLC. In recent years, based on the development of genetic engineering and tumor biology, genetic diagnosis and small molecule targeted drugs have shined in the diagnosis and treatment of various cancers. Therefore, an accurate understanding of the molecular mechanism of the occurrence and development of HCC...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00A61P35/00C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 许秋然涂康生黄东胜胡晓歌陆启亮
Owner ZHEJIANG MEDICAL COLLEGE
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