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Marker (TP63) for predicting occurrence of cervical lesions of HPV (human papilloma virus)-positive patients

A technology of cervical lesions and markers, applied in the field of tumor molecular biology, can solve the problems of subjectivity, lack of quality control, lack of unified authoritative consensus, fluctuations in sensitivity and specificity, etc., achieve low false positive rate and reduce work Quantitative, sensitive diagnosis or the effect of

Pending Publication Date: 2019-10-25
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method of cytological examination depends on the experience and technical level of the examiner to a large extent, is highly subjective and lacks quality control, and the sensitivity and specificity fluctuate greatly. There is no unified and authoritative consensus on the treatment of atypical squamous cells of undetermined significance (ASC-US)

Method used

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  • Marker (TP63) for predicting occurrence of cervical lesions of HPV (human papilloma virus)-positive patients
  • Marker (TP63) for predicting occurrence of cervical lesions of HPV (human papilloma virus)-positive patients
  • Marker (TP63) for predicting occurrence of cervical lesions of HPV (human papilloma virus)-positive patients

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Using immunohistochemical staining to detect the expression of TP63 protein in different tissue samples

[0026] Cases: From November 2016 to December 2017, patients undergoing cervical cancer screening at the First Affiliated Hospital of Sun Yat-sen University. Of the 227 HPV(+) patients, 121 underwent colposcopy, 23 were normal, 24 were CIN I, and 74 were CIN II. Select 10 cases of HPV(+), LCT(-)

[0027] 131 patients were used as controls for immunocytochemical examination, and the case screening process is shown in Figure 1.

[0028] experimental method:

[0029] 1. Immunohistochemical staining was used to detect the expression of TP63 protein in different tissue samples. See Table 1 for the histological information in the histological chips CIN481, CIN1201, CR501 and BC10025.

[0030] (1) Dewaxing and hydration: After baking paraffin sections in a constant temperature oven at 60°C for 2 hours, immerse them in xylene 3 times, 5 minutes each time; 2 ...

Embodiment 2

[0046] Example 2. Immunocytochemical method to detect the expression of TP63 protein in different cell samples.

[0047] (1) Dewaxing and hydration: put the prepared cell smear in a 60°C incubator and bake for 2 hours, then immerse in xylene 3 times, 5 minutes each time; 2 times in absolute ethanol, 5 minutes each time; In 95%, 85%, and 75% ethanol for 5 minutes each; wash with PBS 3 times, 5 minutes each time.

[0048] (2) Eliminate endogenous peroxidase: Remove the liquid around the tissue on the smear, place it flat in a wet box, add 3% hydrogen peroxide solution dropwise, incubate at room temperature for 10-15min, wash with PBS 3 times, 5min each time .

[0049] (3) Antigen retrieval: Put the smear into citrate buffer, put it into a pressure cooker and heat it to boiling, 800w, cover the pressure valve and wait for 1-4 minutes after spraying. The temperature of the buffer was naturally cooled to room temperature, and washed 3 times with PBS (containing Triton X-100), 5 m...

Embodiment 3

[0059] Example 3: Sensitivity and Specificity of TP63 Gene as a Molecular Marker in Human Papillomavirus Positive Patients

[0060] The specificity and sensitivity of TP63 as a molecular marker were identified by comparing the CIN I+ and CIN II+ stages of HPV-positive patients with TP63 single index detection with LCT detection. The results are shown in Table 2 and figure 2 , 3 shown.

[0061] Table 2: Comparison results of the efficacy of TP63 and LCT in detecting CIN I+ and CIN II+

[0062]

[0063] From Table 2 and Image 6 , 7 It can be seen that the sensitivity and specificity of detecting CIN I+ and CIN II+ in HPV positive patients using TP63 single index are higher than those of LCT detection. The area under the ROC curve of TP63 protein detection was higher than the area under the ROC curve of cytology detection (P<0.001), suggesting that the detection efficiency of TP63 protein in predicting follow-up histological positivity was higher than that of LCT detect...

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Abstract

The invention discloses a marker for predicting the occurrence of cervical lesions of HPV (human papilloma virus)-positive patients. The marker is a TP63 gene or a protein encoded thereby, the amino acid sequence of the marker is represented by SEQ NO. 1, and the biomarker has an HPV integration site. The biomarker can assist in the diagnosis of cervical high-grade intraepithelial neoplasia of theHPV-positive patients, or screening of people groups suffering from precancerous lesions and having a risk of developing cervical cancers for focused monitoring. The marker for detecting high-risk populations of the HPV-positive patients has the advantages of rapidness, accuracy, convenience, low false positive rate and the like, and has important significance in clinical diagnosis.

Description

technical field [0001] The invention relates to the field of tumor molecular biology, in particular to the application of a tumor marker in predicting the prognosis of cervical cancer. Background technique [0002] Cervical cancer is one of the most common gynecological malignancies in women worldwide after breast cancer, and its incidence rate ranks second among female malignancies. Its incidence rate ranks first in some developing countries. According to the International Cancer Research Center, there are about 527,600 new cases of cervical cancer each year, accounting for about 7.9% of all new cancer cases in women, 84% of which occur in developing countries; there are about 265,700 cervical cancer deaths each year , accounting for approximately 7.5% of all new cancer cases in women, 87% of which occurred in developing countries. my country has a vast territory and a large population. According to the latest national cancer report released by the National Cancer Center ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574C07K14/47
CPCC07K14/47C12Q1/6886C12Q2600/118C12Q2600/158G01N33/57442G01N33/57484G01N2800/52
Inventor 胡争崔资凤田瑞樊惟文陈怡李金庄
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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