Detection method for regulating Hh signal channel to inhibit HL-60 cells

A signaling pathway and detection method technology, applied in the medical field, can solve problems such as unclear whether the HH signaling pathway has a role, and the impact on the survival of HL-60 cells

Inactive Publication Date: 2017-06-30
SHIHEZI UNIVERSITY
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Problems solved by technology

[0004] The purpose of the present invention is to provide a detection method for regulating the Hh signaling pathway to inhibit HL-60 cells, aiming to solve the problem of whether the HH signaling pathway has an effect on the growth of bone marrow stromal cells to leukemia cells and whether the bone marrow stromal cells induce it. The problem of the effect of HH signaling on the survival of HL-60 cells

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  • Detection method for regulating Hh signal channel to inhibit HL-60 cells
  • Detection method for regulating Hh signal channel to inhibit HL-60 cells
  • Detection method for regulating Hh signal channel to inhibit HL-60 cells

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[0041] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0042] The detection method for regulating the Hh signaling pathway and inhibiting HL-60 cells provided by the embodiments of the present invention includes:

[0043] The kit was used to detect the proliferation of HL-60 cells among different experimental groups; HL-60 cells were inoculated in 96-well plates with or without bone marrow stromal cells HS-5, and the absorbance OD value of each well was measured with a microplate reader, and plotted. Proliferation curve of HL-60 cells;

[0044] AnnexinⅤ-FITC / PI double staining was used to detect the apoptosis rate of HL-60 cells in each group; HL-60 cells were inoculated in 24...

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Abstract

The invention discloses a detection method for regulating an Hh signal channel to inhibit HL-60 cells. The detection method comprises the steps as follows: proliferation of HL-60 cells among different experiment groups is detected with a kit, the HL-60 cell apoptosis rate of each group is detected through double staining, expression of Hedgehog signal channel components, namely, GLI1 genes and apoptosis genes BCL-2 and BCL-XL, of each experiment group is detected with a semi-quantitative RT-PCR method, and expression of GLI1 protein is detected with an immunofluorescence method. A co-culture mode is established in vitro by use of human acute myelocytic leukemia cell strains HL-60 and HS-5 cells, the interaction of in-vivo leukemia cells and hemopoietic microenvironment around the leukemia cells is simulated, the possible mechanism of the effect and action of the hemopoietic microenvironment on leukemia cell apoptosis is researched, and a theoretical foundation is laid for studying leukemia treatment drugs targeted at blocking the interaction of the hemopoietic microenvironment and the leukemia cells.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a detection method for regulating Hh signaling pathway to inhibit HL-60 cells. Background technique [0002] Bone marrow stromal cells (BMSCs) are the core components of the hematopoietic microenvironment (HM). Bone marrow stromal cells in the bone marrow hematopoietic microenvironment can regulate the proliferation, survival and drug resistance of acute myelogenous leukemia (AML). Therefore, in addition to directly targeting the treatment of acute myeloid leukemia, blocking the interaction between leukemia and bone marrow stromal cells will provide a new strategy for the treatment of leukemia. Hedgehog (HH) protein belongs to the secretory protein family, widely expressed in mammals, non-mammals and other species, involved in the regulation of various tumor formation, organ maturation, angiogenesis, stem cell differentiation, immune cells and embryonic development....

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/533G01N33/574C12Q1/68C12Q1/02
CPCG01N33/6893C12Q1/6886C12Q2600/158G01N33/533G01N33/57484
Inventor 魏虹邓磊
Owner SHIHEZI UNIVERSITY
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