Early diagnosis marker for cervical cancer caused by HPV infection based on plasma exosome protein and application of early diagnosis marker

A diagnostic marker and early diagnosis technology, which can be applied to medical preparations, drug combinations, and pharmaceutical formulations containing active ingredients, can solve problems such as adverse effects, patient discomfort, and lack of prognostic judgment. Extraction is convenient and easy to operate

Active Publication Date: 2020-08-25
SHENZHEN HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It will also cause discomfort and psychological adverse effects to patients
[0007] Moreover, the HPV DNA test and TCT test require the tester not to have sex within three days, and it is not suitable for the tester after vaginal cleaning; it is not convenient to perform HPV and T

Method used

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  • Early diagnosis marker for cervical cancer caused by HPV infection based on plasma exosome protein and application of early diagnosis marker
  • Early diagnosis marker for cervical cancer caused by HPV infection based on plasma exosome protein and application of early diagnosis marker
  • Early diagnosis marker for cervical cancer caused by HPV infection based on plasma exosome protein and application of early diagnosis marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1. Correlation between plasma exosome Mortalin expression and cervical cancer stage

[0089] Protein non-labeled quantitative technology (Lable-free) can perform mass spectrometry analysis on protein enzymatic peptides through liquid chromatography-mass spectrometry without specific quantitative labeling reagents. By analyzing mass spectrometry data, the signal intensity of specific peptides between samples is compared, thereby To achieve the purpose of relative quantification of the protein corresponding to the peptide.

[0090] The present invention uses Lable-free to verify the correlation between the expression level of plasma exosome Mortalin and the stage of cervical cancer. Specific steps are as follows:

[0091] (1) Take blood samples from 10 patients in each of the precancerous lesion group (uterine intraepithelial neoplasia, CIN, group A), cancer group (cervical cancer, group B) and healthy control group (group C), and divide the 10 patients The samp...

Embodiment 2

[0101] Example 2. Correlation between HPV infection and up-regulation of exosome mortalin expression

[0102] Cervical cancer cell lines SiHa (HPV+) and C-33A (HPV-) and immortalized cervical epithelial cells H8 were selected to represent HPV-positive (HPV+) cervical cancer cells, HPV-negative (HPV-) cervical cancer cells and precancerous lesion cells, respectively. Caski is also HPV positive. The correlation between the HPV characteristic protein HPV E6 / E7 and the expression of mortalin in cells and exosomes was compared.

[0103] Extraction of intracellular exosomes

[0104] The specific method is as follows:

[0105] (1) Fetal bovine serum (FBS) was diluted with PRIM1640 medium at a volume ratio of 4:1, and centrifuged at 160,000 g for 16 h in an ultrahigh-speed centrifuge to remove serum exosomes. The obtained diluted serum was filtered through a 0.22 μm filter for aseptic treatment. Take 50ml of the above treated diluted serum, mix it with 50ml PRIM1640 medium, and ma...

Embodiment 3

[0135] Example 3 In vitro cell experiment of exosome Mortalin on malignant transformation of immortalized cervical epithelium

[0136] Immortalized cervical epithelial cells H8, as HPV-infected positive cells, are immortalized cells without canceration, and the mechanism of canceration has not yet been elucidated. This part explains that cervical cancer-derived exosome mortalin has a malignant transformation function on H8 cells, suggesting that the effect of exosome mortalin may be one of the mechanisms of H8 carcinogenesis.

[0137] Knockdown Mortalin SiHa (KdMorSiHa / KdMS) and Knockdown Mortalin Caski (KdMorCaski / KdMC) cell lines were constructed using lentiviral Mortalin shRNA. Both SiHa and Caski are cervical cancer cell lines, and H8 cells are immortalized cervical epithelial cells. Both cell lines were confirmed to be successfully constructed in previous experiments, and it was confirmed that the expression of Mortalin in their exosomes was relatively reduced.

[0138] ...

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Abstract

The invention discloses a plasma exosome protein Mortalin taken as an HPV positive cervical cancer diagnosis marker and an application thereof. Differential proteins of plasma exosomes of a cervical cancer tumor group patient, a precancerous lesion patient (CIN) and a healthy person are compared through proteomics, the mortalin expression condition after the HPV key molecule HPVE6/E7 is knocked out is further determined, that exosome mortalin can serve as a molecular marker of cervical cancer is found, preliminary screening of cervical cancer can be conducted when plasma exosome mortalin is detected, and the early definite diagnosis probability of the cervical cancer can be increased.

Description

technical field [0001] The invention relates to the field of molecular diagnostic markers, in particular to a plasma exosome as an early diagnostic marker for cervical cancer caused by HPV infection and its application. Background technique [0002] Cervical cancer is the second most malignant tumor that harms women in my country. Its risk is second only to breast cancer. Its incidence rate ranks fourth among female tumors, and its mortality rate ranks second. It is the main cause of death from cancer-related diseases in women. Cervical cancer seriously reduces women's quality of life and threatens women's life and health. Therefore, early diagnosis and early treatment of cervical cancer and effective control of cervical cancer incidence and mortality have become a top priority. [0003] HPV infection is one of the main causes of cervical cancer, and its correlation rate is as high as 99.7%. Two necessary conditions for HPV infection to induce cervical cancer are high-risk...

Claims

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Application Information

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IPC IPC(8): G01N33/574A61K45/00A61P35/00A61P35/04
CPCG01N33/57411G01N33/57488A61K45/00A61P35/00A61P35/04
Inventor 郭霞肖悦郝轶陈瑾陈晓娜赵宏丽
Owner SHENZHEN HOSPITAL OF SOUTHERN MEDICAL UNIV
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