Human benign phyllodes tumor cell line GLK-1010 and applications thereof

A technology for tumor cells and benign tumors, applied in the field of tumor cells, can solve problems such as limited value, and achieve the effect of short establishment time and stable biological heredity

Active Publication Date: 2020-04-17
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing molecular markers are also of limited value in predicting tumor biological behavior

Method used

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  • Human benign phyllodes tumor cell line GLK-1010 and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0018] The human phyllodes tumor cell line GLK-1010 of the present invention is obtained by the following method:

[0019] (1) Specimen collection and storage methods: After the complete mammary gland phyllodes tumor has been completely removed, the tumor tissue is cut in the middle with a sterile scalpel, and the vigorously proliferating tissue is taken in DMEM medium. The collection of specimens is carried out under the guidance of the surgeon and the pathologist to prevent the impact on the diagnosis of the pathology report; when the specimens are not used for follow-up operations, they should be stored in DMEM medium at 4°C, and the specimens should be used within 24 hours as much as possible .

[0020] (2) Primary culture: the tissue was washed 3 times with PBS, cut mechanically until it was as small as possible, digested with type III collagenase (1mg / ml, Worthington) and added to a 50ml centrifuge tube containing DMEM / F12, protected from light with tinfoil, 37°C, 180 r...

Embodiment 2

[0035] Embodiment 2 detects the expression situation of virus

[0036] Q-PCR (quantitative real-time polymerase chain reaction, referred to as quantitative PCR) was used to detect the expression level of the immortalized gene (HPV) introduced into the cell line GLK-1010, wherein the primer sequences used to amplify the corresponding gene were listed in Table 1 below. Show.

[0037] Table 1 Primer base sequence

[0038] Forward (forward primer) Reverse (reverse primer) GAPDH AGAGCCTCGAGGAGAAGTTCC (SEQ ID NO. 1) ACTAGGGAGTCAAGGACGGG (SEQ ID NO. 2) HPV TAACTTCTGTTCGGGGAGGC (SEQ ID NO. 3) AGGGAGGGAAAAGATCCCGA (SEQ ID NO. 4)

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Abstract

The invention discloses a human benign phyllodes tumor cell line GLK-1010, which is preserved in China Center for Type Culture Collection, wherein the preservation address is Wuhan University, Wuhan,China, and the preservation number is C2018235. The invention also discloses applications of the human benign phyllodes tumor cell line GLK-1010 as a cell model in research of tumor occurrence and development mechanism and in screening of antitumor drugs. The human phyllodes tumor cell line GLK-1010 disclosed by the invention is established from the source of Chinese people, the line establishmenttime is short, the biological heredity is stable, no human benign phyllodes tumor cell line exists on the market at present, and by using the phyllodes tumor cell line as a research model, great helpcan be provided for knowing the pathogenesis of Chinese benign phyllodes tumor patients.

Description

technical field [0001] The invention relates to the technical field of tumor cells, in particular to a human benign phyllodes tumor cell line GLK-1010 and its application. Background technique [0002] Phylodes tumor of the breast is a rare breast tumor, accounting for about 1% of breast tumors. This tumor grows rapidly and often presents as a huge mass; it is divided into benign, borderline, and malignant histologically; even Benign phyllodes tumors can also recur, and malignant phyllodes tumors can also undergo hematogenous metastasis. The therapeutic effects of chemotherapy and radiotherapy on phyllodes tumors are uncertain. The current treatment that can reduce the probability of recurrence and metastasis is extended surgical resection, but even with extended surgical resection, the local recurrence rate of phyllodes tumors is still as high as 8-36% %, the hematogenous metastasis rate of malignant phyllodes tumors is as high as 22%. [0003] Currently, the mechanisms d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02C12R1/91
CPCC12N5/0693G01N33/5011C12N2503/02G01N2500/10Y02A50/30
Inventor 宋尔卫聂燕黄红颜
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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