Family with sequence similarity 84, member B (FAM84B) antisense nucleotide and application thereof

A technology of antisense nucleotides and nucleotide sequences, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, and the use of vectors to introduce foreign genetic materials, etc., can solve problems such as interference, reduce the degree of malignancy, reduce the number, and improve specific effect

Active Publication Date: 2013-09-18
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the poor effect of colorectal cancer chemotherapy, and provides a FAM84B antisense nucleotide sequence, the nucleotide sequence is SEQ ID No.1, this sequence interferes with the expression of FAM84B, thereby inhibiting the growth of colorectal cancer cells

Method used

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  • Family with sequence similarity 84, member B (FAM84B) antisense nucleotide and application thereof
  • Family with sequence similarity 84, member B (FAM84B) antisense nucleotide and application thereof
  • Family with sequence similarity 84, member B (FAM84B) antisense nucleotide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Construction and identification of pSuper-shRNA-FAM84B expression vector

[0025] (1) siRNA sequence design

[0026] Design the 21nt siRNA target sequence for FAM84B according to the siRNA Target Finder software, design and construct the shRNA forward and reverse oligonucleotide sequences corresponding to the gene silencing expression vector pSUPER, the forward oligonucleotide sequence is SEQ ID No.2 (the antisense oligonucleotide sequence containing oligonucleotide sequence SEQ ID No.1 and its SEQ ID No.1), the reverse oligonucleotide sequence is SEQ ID No.3 (containing oligonucleotide sequence SEQ ID No.3) ID No.1 and its antisense oligonucleotide sequence of SEQ ID No.1).

[0027] The sequence of the control siRNA is SEQ ID No.4. The forward oligonucleotide sequence corresponding to this sequence to construct the gene silencing expression vector pSUPER is SEQ ID No.5, and the reverse oligonucleotide sequence is SEQ ID No.6. for subsequent experiments.

...

Embodiment 2 Embodiment 1

[0046] Inhibition of FAM84B expression by the expression vector constructed in Example 2 Example 1

[0047] The successfully constructed pSuper-shRNA-control and pSuper-shRNA-FAM84B expression vectors were transfected into NCI-H716 cells respectively, and NCI-H716 was stably transfected with pSuper-shRNA-FAM84B cell line, and verified by Real-time RT-PCR Effects of FAM84B silencing.

[0048] Transfection: experimental group: pSuper-shRNA-FAM84B group; control group: pSuper-shRNA-control group.

[0049] The above plasmids were transfected into NCI-H716 cells, and the transfection reagent liposome (PEI) was added, and the operation was performed according to the instructions. After 24 hours of transfection, 2.0 μg / ml of puromycin was added for stable selection, and the culture medium containing the same concentration of puromycin was replaced in 2-3 days. After 7 days, the cells in the blank control group were basically dead, while the cells in the transfection group had scatt...

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Abstract

The invention relates to a family with sequence similarity 84, member B (FAM84B) antisense nucleotide. The nucleotide sequence is SEQ ID No.1; and the FAM84B antisense nucleotide also comprises an expression vector containing the nucleotide sequence and host cells of the expression vector. By the FAM84B antisense nucleotide, the proliferation and the invasion ability of colorectal cancer cells are inhibited by inhibiting FAM84B, and finally, the effect of inhibiting the growth of the colorectal cancer cells is reached.

Description

technical field [0001] The invention relates to a FAM84B antisense nucleotide and its application in the preparation of drugs for inhibiting the growth of colorectal cancer cells, in particular to inhibiting the growth of colorectal cancer cells by inhibiting FAM84B, and belongs to the field of biomedicine. Background technique [0002] Family with sequence similarity84, member B, FAM84B (family with sequence similarity84, member B, FAM84B), FAM84B is also known as BCMP101, the gene is highly expressed in a variety of tumors: breast cancer, chronic myelogenous leukemia, esophageal cancer, prostate cancer, colon cancer cancer, melanoma, etc. The phenomenon of co-amplification of this gene and myc was found in colon cancer, and the expression of this gene is located at the plasma membrane where cells contact each other in breast cancer, and interacts with α1-catenin (α1-catenin). Catenin is a family of intracellular glycoproteins with similar structures, and it is a cytoskele...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63A61K48/00A61P35/00
Inventor 傅松滨计薇孙文靖于旸金焰孟祥宁
Owner HARBIN MEDICAL UNIVERSITY
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