Application of FBP1 (fructose-1,6-bisphosphatase 1) gene

A gene and tumor diagnosis technology, applied in the field of medicine and biology, can solve the problem of unclear relationship between tumors and so on

Inactive Publication Date: 2012-05-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

FBP1 gene (Genbank No. NM_000507), the relationship wit

Method used

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  • Application of FBP1 (fructose-1,6-bisphosphatase 1) gene
  • Application of FBP1 (fructose-1,6-bisphosphatase 1) gene
  • Application of FBP1 (fructose-1,6-bisphosphatase 1) gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 RT-PCR experiment to detect the expression of FBP1 gene in tumor cells

[0028] 1. Cell culture

[0029] All cells were cultured in RPMI-1640 medium containing 100 U / ml penicillin, 100 μg / ml streptomycin and 10% FBS at 37°C in 5% CO 2 cultured in an incubator.

[0030] 2. Total RNA extraction kit

[0031] The total RNA of cells was isolated and extracted with Trizol reagent from Invitrogen Company according to the method provided by the manufacturer. The reagent is produced based on one-step extraction with acidic phenols. The utensils and water used for RNA extraction must be RNase-free to ensure an RNase-free environment in the experiment.

[0032] 3. Extraction of total RNA

[0033] Cultivate the cells to the logarithmic growth phase, drain the culture medium, add 1ml Trizol directly, let stand at room temperature for 5 minutes, and collect the cells into a 1.5ml centrifuge tube. After adding chloroform, centrifuge at 4oC to separate layers; transfe...

Embodiment 2

[0047] Example 2 Real-time quantitative PCR experiment to detect the expression of FBP1 gene in tumor tissue

[0048] 1. Tissue Isolation

[0049] All specimens were pathologically confirmed. Once the surgically resected specimens are removed from the body, the tumor lesions and adjacent tissues are quickly excised and stored in liquid nitrogen.

[0050] 2. Total RNA extraction

[0051] Dry-bake the homogenizer at 200oC for 4 hours to remove RNase and cool down; take the tissue out of the liquid nitrogen quickly and grind it into powder; use a spatula to put the tissue into the homogenizer pre-added with TRIzol reagent, and homogenize A few minutes; transfer the homogenized liquid into an RNase-free centrifuge tube, add chloroform, and centrifuge at 4oC to separate layers; transfer the upper aqueous phase to an RNase-free centrifuge tube, add isopropanol, and centrifuge at 4oC Precipitate RNA; wash the pellet twice with 75% ethanol; air-dry and dissolve the pellet in RNas...

Embodiment 3

[0057] Cloning of embodiment 3 FBP1 cDNA and construction of expression vector

[0058] 1. Synthesis of clone-specific cDNA

[0059] The reverse transcription reaction (RT) used the SuperScript III First Synthesis System for RT-PCR kit from Invitrogen, USA, and 2 micrograms of total RNA was used in each reaction. The total reaction volume is 20 microliters: 2 microliters of 10×RT buffer, 5 microliters of total RNA, 1 microliter of OligodT20 primer, 1 microliter of 10mM dNTP, 4 microliters of 25mM MgCl2, 1 microliter of 0.1M DTT, and 1 microliter of RNaseOUT , SuperScript III reverse transcriptase 1 microliter, high-purity deionized water 4 microliters. The reverse transcription reaction was performed using an ABI PCR instrument, and the specific operating parameters were as follows: 10 minutes at 25oC; 120 minutes at 37oC; 5 minutes at 85oC; and cool to 4oC. Store at -20oC for later use. 50oC for 50 minutes, 85oC for 5 minutes, add RNaseH after cooling to 4oC, and incubat...

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Abstract

The invention relates to the field of medicinal biotechnology, particularly to an application of a new cancer suppressor gene FBP1 in the preparation for a medicine for diagnosing, preventing and treating tumour. The cancer suppressor gene is expressed with a low specificity in a tumour cell and a tumour tissue, and has a function of inhibiting the growth of the tumour cell; therefore, the tumours expressing the cancer suppressor gene with a low specificity are selected, and the expression of the cancer suppressor gene in the tumour cell is specifically recovered. A new method is provided for a tumour targeted therapy.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to the application of a new tumor suppressor gene FBP1 gene in the preparation of drugs for tumor diagnosis, prevention or treatment. Background technique [0002] Tumor development is a multi-factor multi-step process, involving the activation of oncogenes and the inactivation of tumor suppressor genes. By detecting the changes of these oncogenes and tumor suppressor genes, the occurrence of tumors can be detected early at the molecular level, thereby creating opportunities for timely intervention and treatment. With the continuous deepening of tumor research, the diagnosis and clinical treatment of tumors have begun to be carried out at the molecular level. Through technologies such as gene chip and proteomics, new tumor-related molecular markers are being continuously developed, and the level of early diagnosis of tumors is increasing day by day. At the same time, on the ba...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K48/00A61P35/00
Inventor 金洪传王娴刘昕
Owner ZHEJIANG UNIV
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