Application of LncRNA XIST and miR-132 combined with LncRNA XIST for inhibiting proliferation, invasion and metastasis of gastric cancer cells

A technology for gastric cancer cells and uses, applied in the directions of DNA/RNA fragments, recombinant DNA technology, medical preparations with non-active ingredients, etc., to achieve the effect of promoting apoptosis and inhibiting proliferation

Inactive Publication Date: 2020-09-11
湖南省科域生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, it has also been reported in the literature that long non-coding RNAs are involved in the invasion, migration and lymph node metastasis ...

Method used

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  • Application of LncRNA XIST and miR-132 combined with LncRNA XIST for inhibiting proliferation, invasion and metastasis of gastric cancer cells
  • Application of LncRNA XIST and miR-132 combined with LncRNA XIST for inhibiting proliferation, invasion and metastasis of gastric cancer cells
  • Application of LncRNA XIST and miR-132 combined with LncRNA XIST for inhibiting proliferation, invasion and metastasis of gastric cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 LncRNA XIST is significantly upregulated in gastric cancer tissues and cell lines

[0083] Through RT-PCR experiments, it was found that the expression of LncRNA XIST in gastric cancer tissues was significantly higher than that in adjacent normal tissues (P figure 1 A); At the same time, in 65 cases of gastric cancer tissue samples, it was found through statistics that the relative expression level of LncRNA XIST in gastric cancer tissues without metastasis (n=20) was higher than that in gastric cancer tissues with metastasis (n=45) The level was also significantly reduced (P=0.001) ( figure 1 B); Compared with GSE-1 in normal gastric tissue epithelial cells, the expression of LncRNA XIS in each gastric cancer cell line was significantly increased, ( figure 1 C), among them, the expression in SGC7901 gastric cancer cells is relatively the highest, so this cell line is subsequently selected for knockdown treatment and related experiments are carried out. Previ...

Embodiment 2

[0084] Example 2 Effect of LncRNA XIST on Biological Function of Gastric Cancer Cells

[0085] In order to evaluate whether LncRNA XIST has an impact on the biological function of gastric cancer cells, we selected the SGC7901 gastric cancer cell line obtained from the above screening and interfered with it. The results showed that, compared with the si-NC group, si-XIST-1 and si-NC The expression of LncRNA XIST was significantly decreased in the XIST-2 group, indicating successful transfection (Fig. 2A).

[0086] Then, the proliferation of gastric cancer cells in each group was detected by EdU, scratch, Transwell, and flow cytometry ( Figure 2B ),migrate( Figure 2C ), invasion ( Figure 2D ) and apoptosis ( Figure 2E ), the results showed that compared with the si-NC group, the cell proliferation ability and migration and invasion ability of the si-XIST-1 group and the si-XIST-2 group were significantly decreased, while the si-XIST-1 group and the si- The apoptosis rate...

Embodiment 3

[0087] Example 3 LncRNA XIST can competitively adsorb miR-132

[0088] Previous literature has shown that LncRNA XIST exists as a mechanism of ceRNA, so we first analyzed the binding site of LncRNA XIST and miR-132 through online prediction software (Figure 3A). Then through dual luciferase reporter gene verification, the results showed ( Figure 3B ), compared with the NC group, the luciferase activity of the miR-132 binding region of Wt-LncRNA XIST was inhibited by miR-132, while the LncRNA XIST mutant was not inhibited, indicating that miR-132 can specifically bind LncRNA XIST. Co-immunoprecipitation experiments showed (Fig. 3C), compared with IgG, the LncRNA XIST and miR-132 bound to Ago2 were significantly increased (P Figure 3D ), compared with MUT-miR-132, the Ago protein bound by WT-miR-132 was significantly increased (P<0.05), indicating that miR-132 and LncRNAXIST can be directly combined.

[0089] We further transfected miR-132 and LncRNA XIST in the cells, and det...

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Abstract

The invention discloses application of long-chain non-coding RNA XIST and miR-132 combined with the long-chain non-coding RNA XIST for inhibiting proliferation, invasion and metastasis of gastric cancer cells. The si-XIST of the long-chain non-coding RNA XIST can silence the LncRNA XIST, so that the adsorption capacity of the LncRNA XIST to miR-132 is reduced, the miR-132 can be combined with thegene of CUL4B, and finally, the expression of the CUL4B is reduced; therefore, the proliferation, invasion, metastasis and tumor formation capabilities of gastric cancer cells are inhibited, and the apoptosis of the gastric cancer cells is promoted. The application of the LncRNA XIST and miR-132 disclosed by the invention can be beneficial to targeted treatment and clinical treatment of gastric cancer.

Description

technical field [0001] The present invention relates to the use of a long-chain non-coding RNA associated with gastric cancer and a microRNA combined with it in inhibiting the proliferation, invasion and metastasis of gastric cancer cells, the long-chain non-coding RNA is LncRNA XIST, and the microRNA is miR -132 belongs to the field of biomedicine, specifically the field of molecular biology technology. Background technique [0002] Gastric cancer (GC) is a common malignant tumor, and its incidence rate ranks first among all kinds of malignant tumors in my country. Since the vast majority of gastric cancers have no obvious symptoms or specific symptoms in the early stage, they are easily ignored, resulting in a low early diagnosis rate of gastric cancer. Once discovered, it has developed to the middle and late stages, resulting in a high case fatality rate, which can reach 75%. There are four main reasons for the occurrence and development of gastric cancer, one is region...

Claims

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Application Information

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IPC IPC(8): A61K31/713A61K47/54A61P35/00A61P35/04C12N15/113
CPCA61K31/713A61K47/549A61P35/00A61P35/04C12N15/113C12N2310/141
Inventor 李丽丽刘佳利黄娅
Owner 湖南省科域生物医药科技有限公司
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