MYC cancer gene inhibition polypeptide and application thereof

A technique for inhibiting polypeptides and oncogenes, applied in the field of polypeptides for the treatment of tumors, to achieve the effects of inhibiting telomerase activity, inhibiting tumor growth, and promoting the expression of MYC protein

Inactive Publication Date: 2017-05-31
罗瑞雪
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no well-developed MYC oncoprotein preparation available for the treatment of tumors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] In Vitro Cell Proliferation Activity Determination of MYC Oncogene Inhibitory Polypeptide

[0017] The MTT colorimetric method was used. Gastric cancer cell line SGC-7901, which grows logarithmically, was mixed with 1.0×10 5 Add it into a 96-well culture plate and culture for 24 hours. Add different concentrations of the experimental drug MYC oncogene inhibitory polypeptide to the experimental wells and positive drug control wells; add the same volume of solvent to the blank group. Set up five duplicate wells in every hole, culture 48h, add MTT in every well, after 4h of action, add DMSO, hatch 30min, measure absorbance A value at 570nm place of microplate reader, according to the formula cell growth inhibition rate=(experimental group absorbance value / Absorbance value of control group-1)×100%. The calculated IC50 of the experimental drug was 31.21nmol / L.

Embodiment 2

[0019] The in vivo effect of the MYC oncogene inhibitory polypeptide was tested with a tumor model.

[0020] To establish a colon cancer tumor model, HT29 cells in the logarithmic growth phase were digested with trypsin at 37°C, the culture medium was collected, and washed with normal saline for 2 to 3 times. Adjust the cell concentration to 2 × 10 with saline 7 per mL, inoculated subcutaneously in the right forelimb armpit of C57BL / 6 nude mice, each 0.1 mL. After 1w~2w, the tumor volume of nude mice grows to 100~200mm 3 , the tumor tissue was dissected and ground under sterile conditions, and prepared into 1×10 7 cells / mL cell suspension, inoculated subcutaneously in the right forelimb armpit of C57BL / 6 nude mice, 0.1 mL each. After 1w~2w, the tumor volume of nude mice grows to 70~100mm 3 Afterwards, the nude mice were randomly divided into 4 groups, 10 in each group. (1) blank group; (2) low-dose MYC oncogene-inhibiting polypeptide group; (3) medium-dose MYC oncogene-in...

Embodiment 3

[0022] Effect of MYC oncogene inhibitory polypeptide on MYC protein expression in gastric cancer cell line SGC-7901: SGC-7901 cells were cultured at 37°C and 5% CO2 with RPMI-1640 medium containing 10% fetal bovine serum (FBS) When cultivating in the box to a confluence of more than 90%, digest the cells with 0.25% trypsin, count, and centrifuge at 3000rpm for 5min to collect the cells; add RPMI 1640 culture medium (5ml) with 10% calf serum to suspend the cells, count the cells, and adjust the cells The concentration is 5*10 6 cells / ml and cultured in 6-well culture plates. In the experiment, a blank control group and MYC oncogene inhibitory polypeptide groups (5, 10, 20 mg / ml) were set up. After adding 600 μl / well of SGC-7901 cell suspension to each group, 600 μl of 1640 culture medium was added to the blank control group, and different concentrations of polypeptides were added to each dose group of MYC oncogene inhibitory polypeptide. Press 20μl / 1×10 5 Add protein extract...

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PUM

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Abstract

The invention relates to the field of medicines, and in particular relates to polypeptide which can inhibit MYC carcinogenic protein and treat tumor. The sequence of the polypeptide is as shown in SEQ ID NO.1. The polypeptide has the application of tumor treatment. The application includes tumor treatment in multiple administration modes, including subcutaneous or intramuscular injection, intravenous injection or intravenous drip, and the like. The polypeptide has the beneficial effects that in-vitro proliferation activity of multiple tumor cells can be inhibited, MYC protein expression in tumor cells can be promoted, telomerase activity of a tumor cell can be inhibited, growth of tumor inside tumor model naked mice can be inhibited, and the survival rate of tumor-bearing mice can be increased. An effect of treating tumor can be achieved.

Description

[0001] Technical field: [0002] The invention relates to the field of medicines, in particular to polypeptides for treating tumors. Background technique [0003] Tumor treatment has always been a hot and difficult clinical issue. Surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy are the five pillars of cancer treatment today. Targeted therapy is currently the most promising treatment. Despite this, finding therapeutic targets on the tumor side has always been a bottleneck hindering tumor-targeted therapy. [0004] Studies have found that the MYC oncogene is closely related to tumorigenesis. MYC oncogene is a strong oncogene, which can encode MYC oncoprotein. The MYC oncogene regulates the expression of about 10,000 genes and microRNAs in cells. MYC affects the genes that regulate glycolysis, glutamine synthesis, glucose transport, mitochondrial protein synthesis, nucleotide synthesis and other processes, changes bioenergy and metabolism, activates ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00A61K38/16A61P35/00
CPCC07K14/00A61K38/00
Inventor 不公告发明人
Owner 罗瑞雪
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