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Mechanism method of miR-920 targeting YAP1 in drug resistance of esophageal squamous carcinoma chemotherapy

A mir-920, 1.mir-920 technology, applied in the field of chemotherapy resistance of esophageal squamous cell carcinoma targeting YAP1 by miR-920, can solve the problem that the epigenetic regulation has not yet been elucidated

Pending Publication Date: 2022-03-01
CANCER CENT OF GUANGZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As the main effector molecule of this signaling pathway, the relationship between YAP1 and stem cells has been studied the most, but the apparent regulation of YAP1 in esophageal cancer has not been elucidated. Therefore, it is proposed that miR-920 targets YAP1 in esophageal squamous cell carcinoma chemotherapy resistance. The mechanism method in to solve this problem

Method used

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  • Mechanism method of miR-920 targeting YAP1 in drug resistance of esophageal squamous carcinoma chemotherapy
  • Mechanism method of miR-920 targeting YAP1 in drug resistance of esophageal squamous carcinoma chemotherapy
  • Mechanism method of miR-920 targeting YAP1 in drug resistance of esophageal squamous carcinoma chemotherapy

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Embodiment 1

[0043] Immunohistochemical technique was used to detect the expression of YAP1 in esophageal squamous cell carcinoma and chemotherapy-resistant tissues, and fluorescence quantitative PCR was used to detect the expression levels of YAP1 mRNA corresponding to patients in chemotherapy-resistant and chemotherapy-sensitive groups; to explore the expression of YAP1 in esophageal squamous cell carcinoma and chemotherapy-resistant tissues The significance of expression, and the relationship between YAP1 and clinicopathological factors; after the virus was packaged with the lentiviral packaging system of Invitrogen Company, and the esophageal cancer EC109 cells stably interfering with YAP1 were established, the expression of YAP1 and the relationship between YAP1 expression and The relationship between chemotherapy sensitivity; use flow cytometry to sort esophageal cancer stem cell population, combine real-time quantitative PCR and Western Blot to detect the expression of YAP1 in esophag...

Embodiment 2

[0045] In embodiment one, add following operation:

[0046] In step 1, YAP1 is located on human chromosome 11q22, which encodes a squamous protein with a molecular weight of about 65,000. A post-synaptic density protein binding motif comprising a ditryptophan (WW) domain, a TID (transcription activator binding) domain, a TAD (transcription activation) domain and a hydroxyl (-COOH) terminus. YAP1 uses its own structure to interact with many proteins, and plays an extremely important role in multiple signal transduction pathways; the role of YAP1 in stem cells is a current research hotspot, and it maintains through the interaction between TEAD binding domains and TEAD family members. Stem cell function, enhance the expression of genes that maintain the pluripotency of stem cells, such as embryonic stem cell-related genes, Octon transcription factor 4 in the determining region, chromosome Y-2 [(Sexdetermining region Y)-Box 2, Sox-2], leukemia inhibitory factor and bone morphogen...

Embodiment 3

[0049] In embodiment two, add following operation:

[0050] In step 2, according to experimental research results, the characteristics of YAP1 as an oncogene include promoting cell proliferation and inhibiting tumor apoptosis, while enhancing cell proliferation can be achieved by interfering with the expression of cell cycle proteins. The role of YAP1 in the occurrence, development and treatment of tumors has received widespread attention, and many signaling pathways in which YAP1 participates have been verified;

[0051] In step 5, miR-920: MicroRNA (miRNA) is a small non-coding endogenous RNA consisting of 18-24 nucleotides, which is combined in a specific order to mediate its cognate messenger RNA (mRNAs) transcription and regulation, thereby affecting the stability of mRNAs and protein translation. A considerable amount of research data shows that miRNA can affect the established growth process of tumors and the tolerance response to chemotherapy drugs by precisely regula...

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Abstract

The invention relates to the technical field of drug resistance mechanism research, and discloses a mechanism method of miR-920 targeting YAP1 in esophageal squamous cell carcinoma chemotherapy drug resistance, detection of esophageal squamous cell carcinoma and expression of YAP1 in chemotherapy drug resistance tissues by an immunohistochemical technology, and detection of a chemotherapy drug resistance group by a fluorescent quantitative PCR method. According to the invention, firstly, a dual-luciferase reporter gene experiment is mainly carried out on a cellular level to verify that miR920 can directly regulate expression of YAP1 in a targeting manner; on the basis, a miR920 overexpressed esophageal cancer cell line is constructed by using a lentivirus expression technology, and the characteristics of tumor stem cells can be reduced by miR920 targeted down-regulation of YAP1 through a microsphere culture experiment and other modes. The method of MTS, Westernblot, flow cytometry and the like is further utilized to clarify that the apoptosis resistance of esophageal cancer stem cells can be reversed by miR-920 targeting YAP1. Finally, the effects of the miR-920 and the YAP1 in esophageal cancer chemotherapy drug resistance generation are further determined through detection of clinical specimens. Therefore, an experimental basis and a theoretical basis are provided for screening of the esophageal cancer drug resistance reversal agent.

Description

technical field [0001] The invention relates to the technical field of drug resistance mechanism research, in particular to the mechanism and method of miR-920 targeting YAP1 in esophageal squamous cell carcinoma chemotherapy resistance. Background technique [0002] Esophageal cancer is one of the most common malignant tumors in the world. Nearly half of the cases diagnosed each year in the world occur in my country. Squamous cell carcinoma occurs frequently, accounting for 97.6%. Its mortality rate in my country is second only to lung cancer, gastric cancer and liver cancer. Due to the special physiological and anatomical site of the esophagus, atypical early symptoms, and the lack of effective methods for the early diagnosis of esophageal cancer, most patients with esophageal cancer are in the middle and late stages when they see a doctor, and the 5-year survival rate is only 10%-15%. Chemotherapy is usually used to treat advanced tumors, but it is almost inevitable that ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12Q1/6841G01N33/574C12N15/867C12Q1/02
CPCC12Q1/6886C12Q1/6851C12Q1/6841G01N33/57407C12N15/86G01N33/5044C12Q2600/158C12Q2600/178C12Q2600/112C12Q2600/118C12Q2600/106C12N2740/15043C12N2531/00C12N2503/02G01N2333/47G01N2500/10G01N2510/00C12Q2531/113C12Q2563/107
Inventor 徐岗
Owner CANCER CENT OF GUANGZHOU MEDICAL UNIV
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