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Long chain non-coding RNA (Ribonucleic Acid) gene and application method thereof

A long-chain non-coding, application method technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of unknown exact mechanism of development and transfer, and achieve far-reaching significance and promotion. Effect

Active Publication Date: 2013-06-12
CENT SOUTH UNIV
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Problems solved by technology

Hepatocellular carcinoma (HCC) is a major type of primary liver cancer. Although new progress has been made in the diagnosis and treatment of hepatocellular carcinoma, there are still many questions about the exact mechanism of its occurrence, development and metastasis. unknown problem

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  • Long chain non-coding RNA (Ribonucleic Acid) gene and application method thereof
  • Long chain non-coding RNA (Ribonucleic Acid) gene and application method thereof
  • Long chain non-coding RNA (Ribonucleic Acid) gene and application method thereof

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Embodiment Construction

[0034] The present invention is further described below in conjunction with specific embodiments, rather than limiting the present invention.

[0035] 1. New lncRNA gene cloning and analysis

[0036] 1. Materials and methods:

[0037] 1.1 Reagents and kits

[0038] Agarose (agrose) and other commonly used biochemical reagents, gel recovery kits were purchased from Shanghai Huashun Bioengineering Co., Ltd., and 1kbladder DNA molecular weight marker was purchased from Invitrogen; use Mini Kit (Qiagen) extraction kit to extract improved quality RNA, SuperScript TM III (Invitrogen) kit reverse transcription of RNA into cDNA.

[0039] 1.2 Next-generation sequencing technology for RNA sequencing

[0040] After extracting total RNA from human liver cancer tissue samples and normal liver tissue samples, it is purified by column to retain more than 200 nt of RNA;

[0041] Then enrich the mRNA, and use the divalent cation high temperature heating method to fragment the mRNA; use the short mRNA f...

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Abstract

The invention relates to a newly cloned full-length sequence of a long chain non-coding RNA (Ribonucleic Acid) gene and an application method thereof. The gene can be detected in a separation sample by methods including a hybrid method or an amplification method; and the gene can be used as a tumor marker, in particular a marker of liver cancer. According to the gene sequence, a real-time quantification PCR (Polymerase Chain Reaction) primer and an in situ hybridization probe are designed and synthesized; and by detecting the expression level of the long chain non-coding RNA in a liver cancer clinical case sample, expression of the long chain non-coding RNA in liver cancer is up-regulated remarkably, and prognosis of patients with liver caner with high expression of long chain non-coding RNA is poorer. According to the gene sequence, a RNA eukaryotic expression vector in a short hairpin structure for closing the expression of the long chain non-coding RNA by RNA interference is designed and synthesized; and the expression of the long chain non-coding RN in a liver cancer cell line A is inhibited successfully by using the vector.

Description

Technical field [0001] The invention belongs to the field of tumor molecular biology, and specifically relates to a newly cloned long chain non-coding RNA gene sequence and an application method thereof. Background technique [0002] Primary liver cancer is a high-incidence tumor in Asia and parts of Africa. Its fatality rate ranks second among malignant tumors in my country, and the five-year survival rate is only about 5%. The prevention and treatment of liver cancer and its pathogenesis have always been in my country's medical science research. Important subject. Hepatocellular carcinoma (HCC) is a major type of primary liver cancer. Although there have been new advances in the diagnosis and treatment of hepatocellular carcinoma, there are still plenty of precise mechanisms for its occurrence, development and metastasis. Unknown problem. [0003] Long non-coding RNA (lncRNA) is a type of non-coding RNA with a transcript length of more than 200 nucleotides. Studies have found th...

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 曾朝阳李桂源熊炜李小玲唐珂魏芳孛昊龚朝建宋亚莉杨倩李夏雨张文玲范松青石磊马健周鸣
Owner CENT SOUTH UNIV
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