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Application of macc1-as1 probe in preparation of diagnostic reagents for predicting clinical prognosis of gastric cancer

A technology of MACC1-AS1 and diagnostic reagents, which is applied in the function and application fields of genes and RNA, and can solve problems that have not yet been reported on the function of lncRNA

Active Publication Date: 2021-02-19
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] MACC1-AS1 (NR_046756) is a long noncoding RNA (long noncoding RNA, lnc RNA) located on the antisense strand of MACC1, and there is no report on the function of this lnc RNA in malignant tumors

Method used

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  • Application of macc1-as1 probe in preparation of diagnostic reagents for predicting clinical prognosis of gastric cancer
  • Application of macc1-as1 probe in preparation of diagnostic reagents for predicting clinical prognosis of gastric cancer
  • Application of macc1-as1 probe in preparation of diagnostic reagents for predicting clinical prognosis of gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1: High expression of MACC1-AS1 is associated with poor prognosis in patients with gastric cancer.

[0105] In situ hybridization (ISH) was used to evaluate the expression of MACC1-AS1 in paraffin tissue samples from 124 patients with gastric cancer. The results showed that the expression of MACC1-AS1 in gastric cancer tissues was significantly higher than that in adjacent tissues ( Figure 1A ). With the increase of the clinical stage of the patients, the staining score of MACC1-AS1 also increased ( Figure 1B ).

[0106] According to the staining depth of MACC1-AS1, all patients were divided into 3 groups, negative group (0-2 points), weak positive group (3-7 points), strong positive group (8-12 points). Among the 124 gastric cancer patients, there were a total of 8 negative patients and 116 positive for MACC1-AS1, of which 64 were weakly positive and 52 were strongly positive. Taking pathological diagnosis as the gold standard, it can be seen that the sens...

Embodiment 2

[0110] Example 2: In vivo experiments confirm that MACC1-AS1 significantly promotes lung metastasis of gastric cancer cells

[0111] Gastric cancer cell line MKN45 overexpressing MACC1-AS1 was used at 5×10 5 The amount was injected into the body of 4-week-old nude mice through the tail vein, and the neck was cut off 6 weeks after the injection, and the lungs were taken out and photographed ( Figure 2A ) and embedded in paraffin, HE staining and immunohistochemical staining were performed to detect MACC1, Ki67 (proliferation index) and 8-OHdG (oxidative stress level index) ( Figure 2B ), the results showed that the number and volume of lung metastases in the MACC1-AS1 overexpression group were larger than that of the empty control group, the expression of MACC1 and Ki67 was higher, and the expression of 8-OHdG was significantly decreased, indicating that MACC1-AS1 promoted gastric cancer cell lung metastases. Metastasis, promotion of gastric cancer cell proliferation, resist...

Embodiment 3

[0112] Example 3: Compared with normal gastric mucosal epithelial cells, MACC1-AS1 is significantly highly expressed in gastric cancer cells.

[0113] According to the NCBI database, it was determined that MACC1-AS1 was located on the antisense strand of the MACC1 gene ( Figure 3A ). By detecting the expression of MACC1-AS1 in normal gastric mucosal cells and multiple gastric cancer cell lines, it can be found that MACC1-AS1 is highly expressed in gastric cancer cells ( Figure 3B ). Two gastric cancer cell lines, BGC803 and MKN45, were selected for subsequent experiments.

[0114] Since the location of lncRNA in the cell can help determine its function, fluorescence in situ hybridization (FISH) and nuclear and cytoplasmic separation were performed to extract RNA for PCR to detect the distribution of MACC1-AS1 in gastric cancer cells. It can be seen that MACC1-AS1 AS1 is mainly distributed in the cytoplasm ( Figure 3C -D). Immunofluorescence staining also found that MAC...

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Abstract

The invention belongs to the field of functions and applications of genes and RNA, and relates to the application of MACC1‑AS1 probes in the preparation of diagnostic reagents for predicting the clinical prognosis of gastric cancer. The present invention also provides a diagnostic kit for predicting the clinical prognosis of gastric cancer, the kit includes a MACC1-AS1 probe. The present invention reveals for the first time that in gastric cancer, MACC1‑AS1 is related to the prognosis of gastric cancer patients, and is positively correlated with the clinical stage and TNM stage of patients; it is the antioxidant substance produced by gastric cancer cells to maintain redox balance and promote glycolysis when gastric cancer cells are stressed without sugar An important molecule for maintaining glucose metabolism, maintaining survival and reducing apoptosis. Therefore, the present invention provides a theoretical basis and a clinical basis for studying factors related to clinical prognosis of patients with gastric cancer and for clinical application of non-coding RNAs.

Description

technical field [0001] The invention belongs to the field of functions and applications of genes and RNA, and relates to the application of MACC1-AS1, in particular to the application of MACC1-AS1 probes in the preparation of diagnostic reagents for predicting the clinical prognosis of gastric cancer. Background technique [0002] Gastric cancer is the second most fatal malignant tumor in the world, and the incidence and mortality of gastric cancer rank among the top three among malignant tumors in China. And because the early symptoms of gastric cancer are not obvious, more than 2 / 3 of gastric cancer patients are already in the advanced stage when they are first diagnosed. At present, there are many kinds of drugs for gastric cancer chemotherapy, such as alkylating agents, antimetabolites and topoisomerase inhibitors. However, the overall response rate (ORR) of traditional chemotherapy regimens for advanced gastric cancer is only ≥ 40%, and the median survival period is 8-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
Inventor 石敏赵洋刘雅静何婉明黄琼
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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