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Application of Notch1, p16<INK4a> and hTERC genes as screening markers for cervical cancer

A p16ink4a, cervical cancer technology, applied in the field of clinical medicine, can solve the problems of excessive cell proliferation, insignificant correlation between the occurrence of cervical cancer, and low reliability.

Pending Publication Date: 2020-01-03
INNER MONGOLIA UNIV FOR THE NATITIES
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Studies have shown that, p16 INK4a The abnormal expression of the gene is significantly related to the occurrence of cervical cancer and the prognosis of treatment, but it is also reported that p16 INK4a The correlation between protein and cervical cancer is not obvious, but it is more related to the prognosis
Therefore, simply using p16 INK4a Protein as a diagnostic and screening index for cervical cancer is controversial and not reliable
[0005] p16 INK4a The gene is a tumor suppressor gene directly involved in the negative feedback regulation of the cell cycle, p16 INK4a Gene inactivation can lead to excessive proliferation of cells, causing cells that are not fully developed in the G1 phase to enter the S phase ahead of time, leading to tumorigenesis

Method used

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  • Application of Notch1, p16&lt;INK4a&gt; and hTERC genes as screening markers for cervical cancer
  • Application of Notch1, p16&lt;INK4a&gt; and hTERC genes as screening markers for cervical cancer
  • Application of Notch1, p16&lt;INK4a&gt; and hTERC genes as screening markers for cervical cancer

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Experimental program
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Effect test

Embodiment 1

[0026] 1. Materials and methods

[0027] 1.1 Paraffin section of cervical lesion tissue and positive determination of HPV16

[0028] From September 2016 to September 2017, patients with cervical diseases who visited the Affiliated Hospital of Inner Mongolia University for Nationalities and underwent pathological examination were selected, and 23 kinds of HPV genotyping detection kits (PCR fluorescent detection kits) provided by Guangzhou Kaipu Biotechnology Co., Ltd. were used. Acupuncture method), strictly follow the instruction manual to extract HPV DNA and perform PCR amplification, and use ABIQuantistudio DX fluorescence quantitative PCR instrument (ABI company, the United States) for HPV genotyping detection, and 226 patients who were positive for HPV16 were detected, of which 45 Cervical cancer, 35 cases of CIN-Ⅲ, 32 cases of CIN-Ⅱ, 38 cases of CIN-Ⅰ, 42 cases of uterine leiomyoma, 34 cases of chronic cervicitis. The age range was 20 to 71 years old, with an average age...

Embodiment 2

[0043] 2. Results

[0044] 2.1 p16 INK4a , Notch1 and hTERC There were significant differences in the positive expression of the gene in each HPV16 positive cervical lesion tissue.

[0045] The detection of HPV16-positive cervical lesion tissue by immunohistochemistry and fluorescence in situ hybridization showed that see Figure 4 , in cervical cancer, CIN-III, CIN-II, CIN-I, uterine leiomyoma, and chronic cervicitis, p16 INK4a The positive expression rates of the proteins were 100.00% (45 / 45), 91.43% (32 / 35), 59.38% (19 / 32), 52.63% (20 / 38), 42.86% (18 / 42), 26.47% ( 9 / 34); the positive expression rates of Notch1 protein were 91.11% (41 / 45), 51.43% (18 / 35), 56.25% (18 / 32), 52.63% (20 / 38), 45.23% (19 / 34 42), 52.94% (18 / 34); hTERC The positive amplification rates of the genes were 100.00% (25 / 25), 83.33% (10 / 12), 33.33% (4 / 12), 0% (0 / 12), 0% (0 / 9), and 0% (0 / 30), the differences were significant, and p16INK4a was the most significant. And in the detection of the same...

Embodiment 3

[0047] 2. Results

[0048] 2.2 p16 INK4a Gene most strongly associated with development and progression of cervical cancer

[0049] p16 INK4a , Notch1 and hTERC The expression of the gene is constantly changing with the occurrence and development of cervical cancer, showing a positive correlation. p16 INK4a , Notch1 In immunohistochemistry, the gene obviously showed an increase in the number of stained cells and the degree of staining in the tissue ( figure 2 A), hTERC gene in fluorescence in situ hybridization showed that the number of cells with gene amplification and the copy number of the gene increased ( figure 2 B). exist figure 2 In A we can also find p16 INK4a It shows high specificity and sensitivity in the occurrence and development of cervical cancer. Compared with HE staining, its expression only occurs in the lesion and its expression degree is distinct in each stage of cervical cancer development. and Notch1 The expression of the gene is not...

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Abstract

The invention belongs to the technical field of clinical medicine, provides application of Notch1, p16<INK4a> and hTERC genes as screening markers for cervical cancer, and relates to molecular markersfor detecting double or triple genes of cervical precancerous lesion and early diagnosis of the cervical cancer. Patients with cervical disease have no significant symptoms in the early stages of onset, and are easily overlooked and fail to be treated in time, so that the risk of the disease and the difficulty of treatment are increased. It is controversial to use p16<INK4a> protein as a diagnostic screening index for the cervical cancer, and the credibility is not high. The invention provides a set of markers, namely, the application of the Notch1, p16<INK4a> and hTERC genes as the screeningmarkers for the cervical cancer, positive correlation exists between p16<INK4a>, Notch1 and hTERC genes and carcinogenesis of HPV16, and significant positive correlation exists between p16<INK4a> andhTERC genes and the carcinogenesis of the HPV16, and the markers can be used for application of molecular marker reagents for detecting double or triple genes of cervical precancerous lesion and early diagnosis or screening of the cervical cancer.

Description

technical field [0001] The invention belongs to the technical field of clinical medicine, and specifically relates to a molecular marker for dual or triple gene detection for cervical precancerous lesions and early diagnosis of cervical cancer, that is, the application of Notch1, p16INK4a, and hTERC genes as markers for cervical cancer screening. Background technique [0002] Cervical cancer is the most common malignant tumor of the female reproductive system. Its incidence rate is second only to breast cancer and ranks second in female malignant tumors, seriously endangering women's lives and health. Cervical cancer and precancerous lesions are a persistent, progressive, multi-factorial and multi-step disease. Human papilloma virus (HPV) infection is the main cause of cervical cancer, and the positive rate of HPV infection in cervical cancer patients is as high as 99%. [0003] HPV is a non-enveloped double-stranded circular DNA virus consisting of 7900 base pairs. The ge...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/118
Inventor 白海花曹亚宁霍文艳翟率宇强欣吴宁津
Owner INNER MONGOLIA UNIV FOR THE NATITIES
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