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Application of DKC1 and HIF-1 alpha in the synergistic treatment of colorectal cancer

A HIF-1, colorectal cancer technology, applied in the field of biomedicine, can solve the problem of unreported synergistic effect of DKC1 and HIF-1α

Pending Publication Date: 2019-10-08
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The synergistic effect of DKC1 and HIF-1α in tumor suppression has not been reported in the prior art

Method used

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  • Application of DKC1 and HIF-1 alpha in the synergistic treatment of colorectal cancer
  • Application of DKC1 and HIF-1 alpha in the synergistic treatment of colorectal cancer
  • Application of DKC1 and HIF-1 alpha in the synergistic treatment of colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Study on the regulatory effect of DKC1 on the expression of HIF-1α and its target genes

[0065] 1. Experimental method

[0066] (1) Cell lines and cell culture conditions

[0067] Colorectal cancer cell lines DLD1 and HCT116 were obtained from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). DLD1 cells were cultured in RPMI-1640 medium, and HCT116 were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / ml penicillin and 100 μg / ml streptomycin. All cell lines were incubated at 37°C, 5% CO 2 cultured in an incubator.

[0068] (2) RNAi and transiently transfected cells

[0069] siRNA against DKC1 and HIF1A (siDKC1, siHIF-1α) and scrambled siRNA (siCtrl) were purchased from Shanghai Gemma Pharmaceutical Technology Co., Ltd. When cells are 30% confluent, use siLentFect TM Lipid reagents (Bio-Rad Laboratories, Hercules, CA, USA) transfected siRNA into cells.

[0070] The siRNA target sequence is ...

Embodiment 2

[0098] Example 2 Studying the Effects of DKC1 and HIF-1α on Tumor Angiogenesis, Tumor Migration and Invasion

[0099] 1. Experimental method

[0100] (1) Stable transfection cell line construction

[0101] DKC1 stable knockout HCT116 cell line and control cell line were constructed using lentivirus packaging DKC1 shRNA vector and corresponding control vector (purchased from Shanghai Gemma Pharmaceutical Technology Co., Ltd., catalog number: D01005 and D01002, respectively). Then, DKC1 stably knocked out HCT116 cells were infected with HIF-1α overexpressing lentivirus (GeneChem, product number: G0150) to establish a HIF-1α stable overexpressing cell line. These target cells were transfected with lentivirus for 48 hours and then selected with 5 μg / ml puromycin (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) for 2 weeks.

[0102] (2) Immunohistochemistry (IHC)

[0103] Paraffin-embedded CRC tissues and corresponding paracancerous tissues were fixed onto 1.5 mm diameter cores...

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Abstract

The invention discloses an application of DKC1 and of HIF-1 alpha in a synergistic treatment of a colorectal cancer. According to the application of DKC1 and of HIF-1 alpha in the synergistic treatment of the colorectal cancer, an experiment shows that the expression of HIF-1 alpha and VEGF is in positive correlation with the expression of the DKC1. The DKC1 inhibits angiogenesis by adjusting an HIF-1 alpha / VEGF signal path and further inhibits tumor migration and invasion, thus the DKC1, the HIF-1 alpha, and the VEGF can all become drug targets for the treatment of the colorectal cancer, theDKC1 and the HIF-1 alpha / VEGF can be simultaneously suppressed, and the synergistic interaction effect is produced for the treatment of the colorectal cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of DKC1 and HIF-1α in synergistic treatment of colorectal cancer. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignancies in the world. The incidence of CRC ranks third among human tumors, and its mortality rate ranks second among malignant tumors. Most CRC patients are already in the advanced stage of initial diagnosis, and the 5-year survival rate is low. Approximately 25% of CRC patients have liver metastases at initial presentation. Data show that approximately 45% of CRC patients eventually die from the disease due to drug resistance and tumor metastasis after conventional treatment, and 80%-90% of patients with CRC fail to undergo radical resection of liver metastases. Therefore, research to find potential biomarkers of CRC is important for early diagnosis and treatment of CRC. [0003] The dyskeratosis congenita 1 (DKC1) gene...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6886C12Q2600/158
Inventor 白津宋军侯平甫史沛聪江涛
Owner XUZHOU MEDICAL UNIV
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