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Application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer

A ring-shaped, gastric cancer technology, applied in the field of molecular biology, can solve the problems of lack of early diagnosis features and poor five-year survival rate of gastric cancer patients, and achieve the effect of promoting proliferation

Active Publication Date: 2022-03-22
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the abnormal expression of multiple genes involved in the occurrence and development of gastric cancer, there is a lack of effective early diagnostic features, and most gastric cancer patients are diagnosed at an advanced stage, which makes the five-year survival rate of gastric cancer patients poor, and the median survival period of patients with gastric cancer metastasis is less than one year

Method used

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  • Application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer
  • Application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer
  • Application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer

Examples

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Effect test

Embodiment 1

[0029] Example 1 Expression of hsa_circ_0004872 in gastric cancer

[0030] 1. hsa_circ_0004872 is a circular RNA formed by reverse splicing of exons 2, 3, and 4 of MAPK1

[0031] We screened the differentially expressed circular RNAs in gastric cancer and corresponding paracancerous tissues by using the next-generation circular RNA sequencing technology, and selected hsa_circ_0004872, which was significantly differentially expressed, as our candidate research molecule. Through qRT-PCR detection of fresh gastric cancer tissues and corresponding paracancerous tissues, it was found that the expression of hsa_circ_0004872 in gastric cancer tissues was significantly down-regulated (85.7% p<0.0001), so we chose hsa_circ_0004872 (see Table 1 for the sequence, sequence number SEQID NO .1) as our research object.

[0032] Table 1 hsa_circ_0004872 sequence list

[0033]

[0034] hsa_circ_0004872 (hsa_circ_27008) comes from the reverse splicing of exons 2, 3, and 4 of the MAPK1 gene...

Embodiment 2

[0042] Example 2 The effect of hsa_circ_0004872 on the proliferation, invasion and metastasis of gastric cancer cells can significantly inhibit the proliferation of gastric cancer cells

[0043] 1. Effects of overexpression of hsa_circ_0004872 on the proliferation, invasion and metastasis of gastric cancer cells

[0044] To study the effect of hsa_circ_0004872 on the biological effects of gastric cancer cells, we constructed the hsa_circ_0004872 overexpression vector pLCDH-circ-4872. It and the control vector were transfected into BGC-823 and SGC-7901 respectively. The results of qRT-PCR showed that after transfection of the overexpression plasmid, the expression level of hsa_circ_0004872 in gastric cancer cells was significantly increased ( figure 2 A).

[0045] After transfecting the hsa_circ_0004872 overexpression plasmid, the cell proliferation efficiency was detected by EdU assay and CCK8 assay, and it was found that overexpression of hsa_circ_0004872 significantly inhi...

Embodiment 3

[0051] Example 3 The molecular mechanism of hsa_circ_0004872 inhibiting the proliferation, invasion and metastasis of gastric cancer cells

[0052] 1. Subcellular localization of hsa_circ_0004872 in gastric cancer cells

[0053] Since the functions of circRNAs are usually related to their different localizations in cells, current studies have shown that circRNAs formed by exons usually exist in the cytoplasm. To detect the localization of hsa_circ_0004872 in gastric cancer cells, we performed fluorescence in situ hybridization (FISH) experiments. We designed the hsa_circ_0004872 probe with Cy3 red fluorescent label (see Table 3 for the sequence). After the gastric cancer cells grew on the cell slides, the cells were fixed and permeabilized, and the Cy3 red fluorescently labeled probe was hybridized with the cell slides overnight, stained with DAPI, sealed, and photographed under a laser confocal microscope. The experimental results show that: hsa_circ_0004872 mainly exists i...

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Abstract

Application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer. The present invention belongs to the field of molecular biology and provides a circular RNA, the nucleotide sequence of the gene of the circular RNA hsa_circ_0004872 is shown in SEQ ID NO.1, and the circular RNA hsa_circ_0004872 is from the MAPK1 gene of chromosome 22 The 2nd, 3rd, and 4th exons of MAPK1 pre-mRNA are reversely cleaved, and the AG base at the 3' end of the 4th exon of MAPK1 pre-mRNA is connected with the CT base at the 5' end of the 2nd exon, forming a covalently closed loop shape RNA. The circular RNA hsa_circ_0004872 can be used as a gastric cancer diagnostic marker in the preparation of gastric cancer diagnostic reagents.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to the application of circular RNA hsa_circ_0004872 in the diagnosis of gastric cancer. Background technique [0002] Gastric cancer is a common malignant tumor of the digestive system. It is one of the fifth most common cancers in the world. Its mortality rate ranks third in the global cancer mortality rate. In 2018 alone, there were more than 1 million new gastric cancer patients, and it is estimated that they died of gastric cancer More than 783,000 patients. The traditional treatment for gastric cancer is surgical treatment, with radiotherapy or chemotherapy as adjuvant treatment. However, due to the abnormal expression of multiple genes involved in the occurrence and development of gastric cancer, there is a lack of effective early diagnostic features, and most gastric cancer patients are diagnosed at an advanced stage, which makes the five-year survival rate of ga...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 刘志方马存英王效莹臧翊辰
Owner SHANDONG UNIV
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