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Application of MiR-199a/miR-218 combined target gene in detection of hexavalent chromium exposure carcinogenesis process

A technology of mir-218 and mir-199a, which is applied in the technical field of biomarker detection and application, can solve the problems of tumor suppressor gene deletion, hypoxia-inducible factor expression up-regulation, oncogene activation and other problems, and achieves the effect of inhibiting cell angiogenesis. Effect

Inactive Publication Date: 2022-05-06
ZHENGZHOU UNIV
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Problems solved by technology

Current research progress has found that aerobic glycolysis can lead to up-regulation of hypoxia-inducible factor 1 (HIF-1), loss of tumor suppressor genes (such as TP53 and VHL), and activation of other oncogenes (such as AKT, MYC, RAS and PI3K )

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  • Application of MiR-199a/miR-218 combined target gene in detection of hexavalent chromium exposure carcinogenesis process
  • Application of MiR-199a/miR-218 combined target gene in detection of hexavalent chromium exposure carcinogenesis process
  • Application of MiR-199a/miR-218 combined target gene in detection of hexavalent chromium exposure carcinogenesis process

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Embodiment 1-2

[0027] We collected blood samples from chromium-exposed people and normal people, and detected the expression levels of miR-199a, IL-8, miR-218, HK2, and LDHA in samples from each patient by qRT-PCR. By comparing the expression levels of blood samples from normal people, it was found that the expression levels of miR-199a and miR-218 were significantly reduced in the blood samples of the chromium-exposed population ( figure 1 A-B), the expression levels of IL-8, HK2 and LDHA in the blood samples of the chromium-exposed population were significantly increased ( figure 2 A-C). The above results indicated that miR-199a, IL-8, miR-218, HK2 and LDHA played an important regulatory role in the carcinogenesis of chromium exposure.

Embodiment 3

[0029] Bronchial epithelial cells are the main cells responsible for Cr(VI) in respiratory organs. We used the immortalized cell line BEAS-2B (B2B) derived from human bronchial epithelium to perform chromium-induced transformation experiments. After treatment with 1.0 μM sodium chromate for 6 months, malignantly transformed cells (Cr-T) were induced to study the molecular mechanism of chronic Cr(VI) exposure carcinogenesis. The expression levels of miR-199a, IL-8, miR-218, HK2 and LDHA in B2B and Cr-T cells were detected by qRT-PCR. The results showed that compared with B2B cells, the expression of miR-199a and miR-218 was significantly lower in Cr-T cells ( image 3 A-B), the expression levels of IL-8, HK2 and LDHA were significantly increased in Cr-T cells ( image 3 C-D).

Embodiment 4

[0031] In order to further study the downstream signaling pathways in cell lines, we detected the protein expression levels of IL-8, HK2 and LDHA and the correlation of downstream signaling pathways in miR-199a / miR-218 and miR-NC stable expression cell lines by western blot experiments protein. Cells with good growth status and confluence of 80%-95% were selected to extract protein, and the protein concentration was determined by BCA kit, and samples were prepared for electrophoresis. In the cell line overexpressing miR-199a, the expression level of IL-8 was significantly decreased, and the protein expression level of HIF-1α was also significantly decreased ( Figure 4 A-B).

[0032] To detect the effect of miR-199a on angiogenesis, fertilized eggs were cultured at 37°C, Cr-T / miR-NC and Cr-T / miR-199a cells were trypsinized and counted in serum-free medium and resuspended. Matrigel was mixed with cell suspension at a ratio of 1:1 and implanted into the chorioallantoic membra...

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Abstract

The invention provides application of miR-199a and miR-218 in chromium exposure carcinogenic detection and a medicine, and belongs to the technical field of biomarker detection application. The invention particularly relates to expression down-regulation of miR-199a and miR-218 in a blood sample of a chromium-exposed crowd. In addition, expression of downstream genes IL-8, HK2 and LDHA in blood samples of chromium exposed people is up-regulated. The invention particularly relates to application of miR-199a and miR-218 in reversing occurrence and development of chromium-induced tumors. Chromium-induced cell proliferation, tumor growth and the like are influenced by changing the expression levels of the miR-199a and the miR-218, which shows that overexpression of the miR-199a and the miR-218 can inhibit tumor growth and reverse occurrence of chromium-induced tumors.

Description

technical field [0001] The present invention relates to the technical field of detection and application of biomarkers, in particular to a method for simultaneously detecting miR-199a and miR-218 and combining downstream target genes IL-8, HK2 and LDHA to evaluate cell malignant transformation and tumor development caused by hexavalent chromium exposure process, and reversed the carcinogenic risk of chromium exposure by a drug carrier containing miR-199a, IL-8, miR-218, HK2, and LDHA. Background technique [0002] As a malignant tumor with the highest morbidity and mortality rate in our country, lung cancer has been widely concerned about its etiology, and its pathogenesis has been continuously explored. Among them, lung cancer induced by environmental factors has also attracted more and more people Among them, chromium (Chromium, Cr) is a widely recognized environmental pollutant and occupational respiratory tumor carcinogen. Chromium is mainly produced in chromium-contain...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00A61P35/00C12Q2600/158C12Q2600/178
Inventor 王琳刘文静江秉华周志豪
Owner ZHENGZHOU UNIV
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