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Application of MYCBP2-AS1 in preparation of products for diagnosis and treatment of cervical cancer

A MYCBP2-AS1, 1.MYCBP2-AS1 technology, which is applied in the directions of medical preparations containing active ingredients, recombinant DNA technology, biochemical equipment and methods, etc.

Inactive Publication Date: 2018-12-18
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the role of lncRNA in the process of cervical carcinogenesis.

Method used

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  • Application of MYCBP2-AS1 in preparation of products for diagnosis and treatment of cervical cancer
  • Application of MYCBP2-AS1 in preparation of products for diagnosis and treatment of cervical cancer
  • Application of MYCBP2-AS1 in preparation of products for diagnosis and treatment of cervical cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 High-throughput sequencing screening of differentially expressed genes

[0035] 1. Sampling

[0036] From November 2013 to November 2015, 9 cases of paracancerous tissue samples from 25 patients with cervical cancer treated in the Department of Obstetrics and Gynecology, China-Japan Friendship Hospital of Jilin University were collected. Store in 80°C low temperature refrigerator.

[0037] 2. Total RNA extraction from tissue samples

[0038] use Reagent (invitrogen, Cat. No. 15596-018) was used to extract sample RNA, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0039] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0040] ① Add Trizol and store at room temperature for 5 minutes;

[0041] ② Add 0.2mL of chloroform, vibrate the centrifuge tube vigorousl...

Embodiment 2

[0050] Example 2 RT-PCR verification of MYCBP2-AS1 expression in cervical cancer tissue

[0051] 1. Materials

[0052] 20 cervical cancer tissue samples and 8 paracancerous tissue samples were obtained from the Department of Obstetrics and Gynecology, China-Japan Friendship Hospital of Jilin University during cervical cancer surgery from November 2013 to November 2015, and they were grouped and numbered. All samples were confirmed by pathological examination. All sample numbers were stored in a -80°C low-temperature refrigerator.

[0053] 2. Method

[0054] 2.1 Extract the total RNA from the tissue sample, the same as the extraction method in Example 1.

[0055] 2.2 Synthesis of cDNA by reverse transcription

[0056] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0057] Using a reverse transcri...

Embodiment 3

[0071] Example 3 RNAi interferes with the expression of MYCBP2-AS1 and its effect on cervical cancer cells

[0072] 1. Materials

[0073] 1. Source of cells

[0074] Human cervical cancer cell C-33A (purchased from ATCC cell bank);

[0075] 2. siRNA design and synthesis

[0076] According to the online design software siDirect version 2.0 (http: / / design.rnai.jp / ), according to the gene sequence chr13:76990659-77327044, design the corresponding siRNA, the specific sequence is shown in Table 3. After the design is sent to the synthesis company for synthesis.

[0077] Table 3 List of siRNA sequences

[0078]

[0079] 2. Experimental method

[0080] 1. Cell grouping

[0081]Group C: blank control group; Group C1: liposome transfection group; C2 group: non-specific siRNA-NC transfection group; S1, S2 groups: specific siRNA transfection group.

[0082] 2. Transfection

[0083] Follow Lipofectamine TM The steps provided by 2000Transfection Reagent were carried out.

[00...

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Abstract

The invention discloses an application of MYCBP2-AS1 in preparation of products for diagnosis and treatment of cervical cancer, and further confirms that the expression of the MYCBP2-AS1 in a cervicalcancer tissue is up-regulated. The invention further discloses the MYCBP2-AS1 for preparing a diagnostic kit for detecting cervical cancer. The kit includes primers specifically amplifying cervical cancer-related lncRNA and instructions, wherein the cervical cancer-related lncRNA is the MYCBP2-AS1. The invention also discloses an MYCBP2-AS1 inhibitor, wherein the inhibitor includes siRNA of MYCBP2-AS1. The MYCBP2-AS1 is used for detecting cervical cancer, not only can early detection be quickly and effectively achieved, but also therapeutic targets and important basis are provided for clinical applications such as gene therapy and drug therapy.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of MYCBP2-AS1 in the preparation of products for diagnosing or treating cervical cancer. Background technique [0002] Cervical cancer is a malignant tumor that appears at the junction of endocervical epithelial cells and squamous epithelial cells in the transition zone of the cervix or vagina. Cervical cancer ranks second in female incidence after breast cancer, and its mortality rate ranks first among gynecological malignancies. It is one of the most common malignant tumors that threaten women's health and life. In recent years, due to factors such as environmental pollution, increased social work pressure, and poor hygiene habits in life, the incidence of cervical cancer has increased significantly, and the trend of younger patients is becoming more and more obvious. [0003] The occurrence, development and prognosis of cervical cancer is a complex multi-stage proce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113A61K31/7088A61P35/00A61P15/14
CPCA61P15/14A61P35/00C12N15/113C12N2310/113C12N2310/14C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 王俊容刘大海胡宇博叶聪
Owner JILIN UNIV
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