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Journal of Traditional Chinese Medicine >

2024 , Vol. 44 >Issue 4: 703 - 712

DOI: https://doi.org/10.19852/j.cnki.jtcm.20240617.002

Original articles

High-throughput sequencing analysis of differential microRNA expression in the process of blocking the progression of chronic atrophic gastritis to gastric cancer by Xianglian Huazhuo formula (香连化浊方)

  • Yuxi GUO ,
  • Ze LI ,
  • Nan CHENG ,
  • Xuemei JIA ,
  • Jie WANG ,
  • Hongyu MA ,
  • Runyuan ZHAO ,
  • Bolin LI ,
  • Yucong XUE ,
  • Yanru CAI ,
  • Qian YANG
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  • 1 Department of spleen and stomach diseases, First Affiliated Hospital of Hebei University of Traditional Chinese Medicine, Shijiazhuang 050000, China
    2 Department of Traditional Chinese Medicine, Hebei General Hospital, Shijiazhuang 050051, China
    3 Department of Gastroenterology, the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, China
    4 College of Integrative Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050011, China

Received date: 2023-03-22

  Accepted date: 2023-07-24

  Online published: 2024-06-17

Supported by

Construction Project of National Clinical Research Base of Traditional Chinese Medicine(Science Letter [2018] No. 131, State Office of Traditional Chinese Medicine);Natural Science Foundation of Hebei Province: Study on the Mechanism of Action of Traditional Chinese Medicine on Disease and Syndrome(H2023423001);Key Research Project of the Ministry of Science and Technology(2018YFC1704100);Key Research Project of the Ministry of Science and Technology: Li Diangui Famous Old Chinese Medicine of Traditional Chinese Medicine Academic View Characteristic, Diagnosis and Treatment Methods and Experience of Prevention and Control of Major Diseases(2018YFC1704102);Provincial Science and Technology Program of Hebei Province: Prevention and Treatment of Gastric Cancer by Blocking the "Inflammation-Cancer Transformation" Based on the Theory of Turbidimetric Toxicity(21377724D);Provincial Science and Technology Program of Hebei Province: to Study the Clinical Efficacy and Mechanism of Huazhuo Jiedu Formula in the Treatment of Chronic Atrophic Gastritis based on Epidermal Growth Factor Receptor/Mitogen Activated Protein Kinase/Extracellular Signal-Regulated Kinase Signaling Pathway(21377740D);Scientific Research Project of Hebei Administration of Traditional Chinese Medicine: Clinical Study of Huazhuo Jiedu Formula Blocking the Pathological Evolution of Chronic Atrophic Gastritis(2022026);Scientific Research Project of Hebei Administration of Traditional Chinese Medicine: Study on the Medication Rules of Spleen and Stomach Diseases of Famous Yanzhao Medical Doctors Based on Data Mining(2022032);Scientific Research Project of Hebei Administration of Traditional Chinese Medicine: to Explore the Mechanism of Xianglian Huazhuo Formula in the Treatment of Chronic Atrophic Gastritis based on Transcriptomics(2023022)

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Abstract

OBJECTIVE: To explore the mechanism of Xianglian Huazhuo formula (香连化浊方, XLHZ) blocking the development of chronic atrophic gastritis (CAG) to gastric cancer (GC) through bioinformatics analysis and in vitro.

METHODS: Pathological morphology of gastric mucosa of rats were observed. High-throughput sequencing was used to analyze the miRNA expression profile of gastric mucosa. The miRanda, miRDB and miRWalk databases were used to predict the differential target genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed for differential target genes. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to verify the differentially expressed miRNAs and target genes. Western blot, EdU, wound healing and flow cytometry were used to observe the effect of XLHZ on epithelial-mesenchymal transition (EMT) markers, proliferation, migration, apoptosis and cell cycle of CAG cells in vitro.

RESULTS: A total of five differentially expressed miRNAs and four differential target genes were screened in this study. GO analysis showed that the target genes were enriched in regulation of neuron development, regulation of transcription factor activity and regulation of RNA polymerase. KEGG pathways database differences in gene enrichment of target genes in the Wnt signaling pathway, Phospholipase D signaling pathway and mitogen-activated protein kinase signaling pathway. qRT-PCR confirmed that miRNAs and its target genes were consistent with the screening results. In vitro, our study revealed that XLHZ could increase the expression of E-cadherin, decrease the expression of transforming growth factor β1, vimentin and β-catenin, inhibite the proliferation and migration of CAG cells, cause cell cycle arrest at G0/G1 and G2/M phase, induce the apoptosis of CAG cells, and prevent the progression of CAG to GC.

CONCLUSION: This study provided a new idea for the mechanism of blocking the progression of CAG to GC by XLHZ, which may be related to the expression of miR-20a-3p, miR-320-3p, miR-34b-5p, miR-483-3p and miR-883-3p and their target genes transferrin receptor, nuclear receptor subfamily 4 member 2, delta like canonical Notch ligand 1 and a kinase anchor protein 12 in CAG. In the future, we will continue to investigate the linkage between the active ingredients of XLHZ and the relevant miRNAs and their target genes, so as to provide more sufficient experimental basis for clinically effective prevention of CAG to GC.

Key words： high-throughput nucleotide sequencing; gastritis, atrophic; stomach neoplasms; microRNAs; Xianglian Huazhuo formula

Cite this article

Yuxi GUO , Ze LI , Nan CHENG , Xuemei JIA , Jie WANG , Hongyu MA , Runyuan ZHAO , Bolin LI , Yucong XUE , Yanru CAI , Qian YANG . High-throughput sequencing analysis of differential microRNA expression in the process of blocking the progression of chronic atrophic gastritis to gastric cancer by Xianglian Huazhuo formula (香连化浊方)[J]. Journal of Traditional Chinese Medicine, 2024 , 44(4) : 703 -712 . DOI: 10.19852/j.cnki.jtcm.20240617.002

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