Method for detecting endometrial receptivity through MST1 and phosphorylated MST1

A technology of MST1 and endometrium, applied in the field of obstetrics and gynecology, can solve the problems of inability to accurately and effectively predict the receptivity of abnormal endometrium

Active Publication Date: 2016-10-05
江苏华朵生物科技有限公司
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Problems solved by technology

[0007] The purpose of the present invention is to provide a method for detecting endometrial receptivity using MST1 and phosphorylated MST1, so as to solve the problem in the prior art that the receptivity of abnormal endometrium cannot be accurately and effectively predicted

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  • Method for detecting endometrial receptivity through MST1 and phosphorylated MST1
  • Method for detecting endometrial receptivity through MST1 and phosphorylated MST1
  • Method for detecting endometrial receptivity through MST1 and phosphorylated MST1

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Embodiment Construction

[0040] The present invention will be further described below in combination with specific embodiments.

[0041] A method for detecting phosphorylated HOXA10 as endometrial receptivity, comprising the following steps:

[0042] (1) Cell culture

[0043] Human endometrial cancer cell lines Ishikawa and HEK293T cells were maintained in DMEM / F12 medium (GibcoBRL / Invitrogen) containing 10% FBS and penicillin and streptomycin (100IU / mL penicillin and 100μg / mL streptomycin) , when the cells grew to 90% confluence, they were routinely digested and passaged with trypsin cell digestion solution (Trypsin-EDTA, Gibco BRL / Invitrogen) with a mass volume ratio of 0.25%, at 37°C, 5% CO 2 , Cultivated in saturated humidity.

[0044] (2) Western Blot

[0045] After Ishikawa or HEK293T cells were washed twice with pre-cooled PBS, 500 μL of cell lysis buffer [50.0mmol / L Tris pH=7.6, 150.0mmol / L NaCl, 0.1% SDS, 1.0% NP-40, Protease Inhibitor Cocktail, Phosphatase Inhibitor Cocktail (Sigma)], th...

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Abstract

The invention provides a method for detecting endometrial receptivity through MST1 and phosphorylated MST1. The method comprises the steps that a human endometrial carcinoma cell line Ishikawa and HEK293T cells are kept and cultured in a 10% FBS and mycillin containing double-antibody DMEM/F12 culture solution; an immunoblotting experiment is carried out; co-immunoprecipitation is carried out; immunohistochemistry is carried out; adjustment of MST1 on the transcriptional activity of HOXA10 is analyzed through chromosome co-immunoprecipitation (ChIP)/PCRs; human choriocarcinoma cell (BeWo) sphere adhering experiments are carried out. According to the method, the expression level of MST1 and MST1 phosphorylation modification in endometrial tissue is detected conveniently and fast, and the receptivity state of the endometrium can be directly reflected.

Description

technical field [0001] The invention belongs to the field of obstetrics and gynecology, in particular to a method for detecting endometrial receptivity. Background technique [0002] The successful implantation of embryos is inseparable from the receptive and well-developed maternal inner membrane. In the late period of the menstrual cycle, affected by the estrogen and progesterone secreted by the ovary, the normal endometrium undergoes a series of regular changes in morphology and biochemistry, develops and forms a receptive endometrium, and prepares for embryo implantation. However, in the IVF-ET cycle, endometrial receptivity developmental defects in patients with adenomyosis, thin endometrium, and unexplained repeated implantation failures cause embryo implantation failures, which also restrict the further improvement of the clinical pregnancy rate of IVF-ET. important reason. At the same time, as the third plenary session of the 18th National Congress of the Communist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/573
Inventor 颜桂军孙海翔蒋玥闫蔷程茜
Owner 江苏华朵生物科技有限公司
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