Application of dendrophenol in preparing medicines for inhibiting proliferation of cervical carcinoma cells and for inducing autophagic apoptosis of cervical carcinoma cells

A technology for cervical cancer cells and cervix, applied in the field of dendrobium phenol, can solve problems such as cervical cancer that has not yet been seen, and achieve the effects of inducing cell cycle arrest and low production cost

Inactive Publication Date: 2016-09-21
HEFEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] So far, there has been no report on the use of dendrobium as an anticancer drug in the treatment of cervical cancer

Method used

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  • Application of dendrophenol in preparing medicines for inhibiting proliferation of cervical carcinoma cells and for inducing autophagic apoptosis of cervical carcinoma cells
  • Application of dendrophenol in preparing medicines for inhibiting proliferation of cervical carcinoma cells and for inducing autophagic apoptosis of cervical carcinoma cells
  • Application of dendrophenol in preparing medicines for inhibiting proliferation of cervical carcinoma cells and for inducing autophagic apoptosis of cervical carcinoma cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Culture of Cells

[0050] 1. Recovery of cells

[0051] (1) Put sterilized pipette tips (10μL, 200μL, 1mL, 5mL) and centrifuge tubes, calf serum (WISENT INC), DMEM culture medium (WISENT INC), UV Sterilization is greater than 2h. Wear laboratory overalls and disposable rubber gloves during operation.

[0052] (2) Prepare hot water at 37°C in advance (38°C in winter), and take out the frozen human cervical cancer Hela-LC 3 cells (cryopreservation tube) from the ultra-low temperature refrigerator (gifted by Professor Wen Longping, School of Life Sciences, University of Science and Technology of China) immediately. Throw it into a water bath and stir it continuously with a glass rod to make it quickly pass through 0°C. After it is completely melted, spray it with alcohol and put it on the ultra-clean workbench.

[0053] (3) Both the serum and the culture medium were overheated, and the serum was added to DMEM at a volume ratio of 1 / 9 to make a cell culture me...

Embodiment 2

[0062] Embodiment 2: MTT detects cell proliferation

[0063] 1. Cell plating

[0064] (1) Prepare 96-well cell culture plates and hemocytometers, and sterilize the required consumables in advance.

[0065] (2) Microscopically examine Hela cells, and digest the cells to make a single cell suspension when they grow to a single layer.

[0066] (3) Count with a hemocytometer, and a bright dot observed under a microscope is a cell; record the number of cells in the eight counting areas as N 1 , N 2 ~N 8 , then the number of cells per milliliter in the suspension is calculated as N=[(N 1 +N 2 +…+N 8 ) / 8]×10 4 . 100 μL / well of Hela cells, 5×10 4 / mL of cells to plate.

[0067] (4) Take an appropriate amount of cell suspension and dilute it into a plating suspension with the required density, and the remaining suspension is used for subculture.

[0068] After plating is complete, mark the cell name and seeding time.

[0069] (5) Put it back into the CO2 incubator, and shak...

Embodiment 3

[0092] Example 3: Cell cycle detection

[0093] 1. Cell sample preparation

[0094] (1) Prepare centrifuge tube (2mL), PBS, trypsin (EDTA-free, 0.25%), and pre-cooled 70% ethanol solution in advance;

[0095] (2) Hela cells after the action of dendrobium, take out the six-well plate marked for 24h, and mark C (1, 2, 3), 50 (4, 5, 6), 100 (7 , 8, 9), 200μmol / L (10, 11, 12);

[0096] (3) Aspirate the cell culture solution in the well and place it in 12 centrifuge tubes (2mL) prepared in advance, and wash the cells once with PBS; , EDTA-free) digested cells;

[0097] (4) Discard the trypsin, add the collected cell culture solution back to the corresponding well to stop the digestion and blow it into a single cell suspension, and transfer it back to the corresponding centrifuge tube again;

[0098] (5) Centrifuge for 5min (1000rpm), discard the supernatant, resuspend in PBS, centrifuge again (1000rpm, 5min), discard the supernatant.

[0099] (6) Take pre-cooled 70% ethanol, r...

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PUM

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Abstract

The invention discloses an application of dendrophenol in preparing medicines for inhibiting proliferation of cervical carcinoma cells and for inducing autophagic apoptosis of cervical carcinoma cells, and belongs to the field of medicines. By inhibiting the proliferation of the cervical carcinoma cells and inducing the autophagic apoptosis activity of the cervical carcinoma cells in vitro, an anti-tumor effect of the dendrophenol in resisting the cervical carcinoma cells is detected, and an anti-tumor concentration of the dendrophenol is analyzed, proving that the dendrophenol has an obvious inhibitory effect on the cervical carcinoma cells; therefore, novel medicines for treating human cervical carcinoma can be developed conveniently. The human cervical carcinoma cells are Hela cells. The effective concentration range of the dendrophenol of inhibiting the proliferation of the cervical carcinoma cells and inducing the autophagic apoptosis of the cervical carcinoma cells is 25-200 [mu]mol/L.

Description

technical field [0001] The invention belongs to the field of biomedicine, and particularly relates to the application of dendrobium in the preparation of drugs for inhibiting the proliferation of cervical cancer cells and inducing autophagy and apoptosis of cervical cancer cells. Background technique [0002] For humans, especially women, cervical cancer is a malignant tumor with an incidence rate second only to breast cancer, and it ranks second in cancer mortality among women. The causes of the disease include unhealthy sex life, premature childbearing age, contempt for thought, unbalanced nutritional elements, smoking, and viral infection (such as HPV, etc.). [0003] The current examination and diagnosis methods for cervical cancer include cytological diagnosis, virus detection and detection of specific markers. Clinically, radiotherapy, chemotherapy, surgery, drugs and other methods are used for treatment. At present, gene therapy is still on the rise. In recent years...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/09A61P35/00
CPCA61K31/09
Inventor 王伟徐忠东陈金武耿明董姗姗
Owner HEFEI NORMAL UNIV
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