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Application of human IGFL3 gene and related product

A technology of genes and uses, applied in medical preparations containing active ingredients, genetic engineering, plant genetic improvement, etc., can solve problems such as the absence of IGFL3 gene, and achieve the effects of inhibiting proliferation, controlling the growth process, and inhibiting the growth of gastric cancer.

Pending Publication Date: 2022-01-14
SHANGHAI GENECHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, there is no relevant report on the use of IGFL3 gene in the treatment of gastric cancer

Method used

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  • Application of human IGFL3 gene and related product
  • Application of human IGFL3 gene and related product
  • Application of human IGFL3 gene and related product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1 Preparation of RNAi lentivirus against human IGFL3 gene

[0098] 1. Screening for effective siRNA targets against the human IGFL3 gene

[0099] Retrieve IGFL3 (NM_207393) gene information from Genbank; design effective siRNA targets for IGFL3 gene. Table 1-1 lists the screened effective siRNA target sequences against the IGFL3 gene.

[0100] Table 1-1 is targeted at the siRNA target sequence of human IGFL3 gene

[0101] SEQ ID NO TargetSeq(5'-3') 1 TGAGGGTTCTGGGTATGAA 2 GTGGGAACAAGATCTACAA

[0102] 2. Preparation of lentiviral vector

[0103] Synthesize double-stranded DNA Oligo sequences (Table 1-2) containing Age I and EcoR I restriction sites at both ends for siRNA targets (taking SEQ ID NO: 1 and 2 as examples); restrict with Age I and EcoR I The endonuclease acted on the pGCSIL-GFP vector (provided by Shanghai Jikai Gene Chemical Technology Co., Ltd.) to make it linear, and the digested fragment was identified by agarose ge...

Embodiment 2

[0122] Example 2 Real-time fluorescent quantitative RT-PCR method to detect gene silencing efficiency

[0123] Human gastric cancer AGS cells in the logarithmic growth phase were digested with trypsin to make a cell suspension (the number of cells was about 5×10 4 / ml) were inoculated in a 6-well plate and cultured until the cell confluency reached about 30%. According to the multiplicity of infection value (MOI, AGS: 20), an appropriate amount of the lentivirus prepared in Example 1 was added, the culture medium was replaced after 24 hours of culture, and the cells were collected after the infection time reached 5 days. Total RNA was extracted according to the instruction manual of Invitrogen's Trizol. According to the M-MLV instruction manual of Promega Company, RNA was reverse-transcribed to obtain cDNA (see Table 2-1 for the reverse transcription reaction system, react at 42°C for 1 hour, and then bathe in a water bath at 70°C for 10 minutes to inactivate the reverse tran...

Embodiment 3

[0130] Example 3 Detection of proliferation ability of tumor cells infected with IGFL3-shRNA lentivirus

[0131] Human gastric cancer AGS cells in the logarithmic growth phase were digested with trypsin to make a cell suspension (the number of cells was about 5×10 4 / ml) were inoculated in a 6-well plate and cultured until the cell confluency reached about 30%. According to the multiplicity of infection (MOI, AGS: 20), an appropriate amount of virus was added, and the culture medium was replaced after 24 hours of culture. After the infection time reached 5 days, the cells of each experimental group in the logarithmic growth phase were collected. The complete medium was resuspended into a cell suspension (2×10 4 / ml), inoculate a 96-well plate at a cell density of about 2000 / well. 5 replicate wells in each group, 100 μl per well. After laying the board, place at 37°C, 5% CO 2 Incubator cultivation. From the second day after plating, the plate was detected and read once a d...

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Abstract

The invention belongs to the field of biological medicine research, and particularly relates to application of a human IGFL3 gene serving as a target to preparation of gastric cancer treatment drugs or gastric cancer diagnosis drugs. Through extensive and deep research, proliferation of gastric cancer cells can be effectively inhibited after expression of the human IGFL3 gene is down-regulated by adopting an RNAi method; and the growth process of gastric cancer can be effectively controlled. siRNA or a nucleic acid construct containing the siRNA sequence and a lentivirus provided by the invention can specifically inhibit the proliferation rate of the gastric cancer cells and inhibit the growth of the gastric cancer, so that the gastric cancer is treated, and a new direction is opened up for the treatment of the gastric cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine research, and specifically relates to the use of human IGFL3 gene and related products. Background technique [0002] IGFL3 (Insulin growth factor-like family member 3, IGF-like family member 3) is a protein-coding gene. Diseases associated with IGF-3 include unilateral or bilateral testicular disease and cryptorchidism. IGFL2 is an important accessory gene of this gene. [0003] So far, there is no relevant report about the use of IGFL3 gene in the treatment of gastric cancer. Contents of the invention [0004] In order to overcome the problems in the prior art, the purpose of the present invention is to provide the use of human IGFL3 gene and related products. [0005] In order to achieve the above object and other related objects, the present invention adopts the following technical solutions: [0006] The first aspect of the present invention provides the use of the human IGFL3 gene as a targe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867C12Q1/6886A61K45/00A61K31/713A61P35/00
CPCC12N15/113C12N15/86C12Q1/6886A61K45/00A61P35/00C12Q2600/136C12Q2600/158C12N2310/14C12N2740/15043C12N2800/107C12N2310/531
Inventor 蔡丽丽曹跃琼
Owner SHANGHAI GENECHEM
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