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A strain of Serratia marcescens and its application in tumor suppression

A kind of technology of Serratia marcescens, Serratia

Active Publication Date: 2019-12-10
蔡剑前 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2002, He Xiaowen used the modified Telomere Repeat Amplification (TRAP)-silver staining method to detect the telomerase activity of Serratia marcescens preparations in 20 cases of bladder transitional cell carcinoma tumor tissues before and after perfusion, and found that Serratia marcescens The preparation can reduce the activity of telomerase in tumor tissue
[0005] In short, although predecessors have done a lot of research on Serratia marcescens and published a number of patent materials, they have not developed into a real clinical drug because of the insufficient efficacy and have remained in the laboratory research stage.

Method used

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  • A strain of Serratia marcescens and its application in tumor suppression
  • A strain of Serratia marcescens and its application in tumor suppression
  • A strain of Serratia marcescens and its application in tumor suppression

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 strain mutagenesis

[0030] (1) strain mutagenesis method

[0031] 1. Strain expansion

[0032] The strain is derived from Serratia marcescens preserved in the China General Microorganism Culture Collection Management Center, and the preservation number is CGMCC1.0589. Under sterile conditions, the original strains were diluted 10-fold and spread on LB slant medium for 24 hours at 37°C. Select a plate with about 20 colonies, pick a single colony with a large, clear boundary, and a milky white color that grows well as the starting strain for ultraviolet mutagenesis, and take its bacterial body in a triangular flask with normal saline and glass beads to make a sterile Bacteria suspension 500mL, 30 ℃, 250rpm water bath 4h, prepared into 10 6 CFU / mL of the mutagenized bacterial suspension, for later use.

[0033] 2. Strain mutagenesis

[0034] Preheat the UV lamp for 25 minutes, take 10 mL of the bacterial suspension and place them on multiple plates (D=9 cm...

Embodiment 2

[0057] Example 2 Preparation of Serratia marcescens bacterin injection

[0058] The preparation method of Serratia marcescens injection is as follows:

[0059] (1) Slant-transfer strains: Serratia marcescens (strain SM-1 screened in Example 1) was inoculated on LB agar medium (pH 7.2) in a Kerner flask, and cultured at 37° C. for 21 hours. Take it out and store it in a -80°C refrigerator for later use.

[0060] (2) Cultivation and fermentation: Take 3 single colonies from the original slant strain of Serratia marcescens, mix them evenly, inoculate them on the LB liquid medium in the Erlenmeyer flask, and cultivate and ferment. The conditions are: LB liquid medium pH 7.2, 10% inoculum size, 75% aeration, temperature 37° C., 21 h, rotation speed 180 rpm.

[0061] (3) Bacteria collection, sterilizing and washing: before collecting bacteria, carry out the pure species inspection such as the morphological characteristics of Serratia marcescens, centrifuge at 4000-6000rpm for 30mi...

Embodiment 3

[0063] Example 3 strain screening

[0064] Observation method of anti-tumor effect was adopted. Six strains of bacteria (SM1, SM2, SM3, SM4, SM5, SM6) were used to treat the mouse tumor-bearing model, and the original strain SM0, the positive drug docetaxel, and Streptococcus A preparation were used as controls, and the tumor tissue size, ascites The formation situation was used as an observation index to determine the strain with the most outstanding anti-tumor effect among the 6 strains.

[0065] 1. Observation of the inhibitory effect of the strain on S180 ascites tumor

[0066] 1.1 Experimental method

[0067] Animal grouping:

[0068] 110 Kunming mice were randomly divided into 11 groups, 10 in each group, respectively normal control group, model control group, SM0 control group, SM1 experimental group, SM2 experimental group, SM3 experimental group, SM4 experimental group, SM5 experimental group , SM6 experimental group, positive drug docetaxel control group, positiv...

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Abstract

The invention discloses a strain of Serratia marcescens, which is classified as Serratia sp. and has been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee on January 11, 2016, and the preservation number is CGMCC NO.11987. The Serratia marcescens of the present invention has an obvious inhibitory effect on A549 non-small cell lung cancer and S180 ascites tumor, and the curative effect is very outstanding. Therefore, the Serratia marcescens can be used as a raw material to prepare a drug for inhibiting tumors, For example: culture the bacteria to obtain a culture, and add or not add other drugs, auxiliary materials, etc. to process and prepare the bacterial agent. The invention provides an excellent strain resource for studying the tumor suppressing mechanism, application, drug development and the like of the Serratia bacterium.

Description

technical field [0001] The invention relates to a strain of Serratia marcescens and its application in inhibiting tumors, belonging to the field of biotechnology. Background technique [0002] According to WHO's "Global Cancer Report 2014", the number of cancer patients and deaths worldwide increased in 2012, and nearly half of the new cases occurred in Asia, most of which were in China and India. Among the four types of malignant tumors including liver, esophagus, stomach and lung, China ranks first in the world in terms of new cases and deaths. Surgical treatment is effective, but most patients have missed the opportunity of surgery when they see a doctor, and can only rely on chemotherapy and radiotherapy, with poor curative effect. Finding therapeutics with new mechanisms is a relentless pursuit in the field. In the 1980s, it was discovered that people infected with bacteria had a lower incidence of skin cancer, lymphoma and leukemia, and found that the injection of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/74A61P35/00C12R1/43
CPCA61K2035/11C12N1/205C12R2001/43
Inventor 蔡剑前张忆军孟红陈雪梅孙永庆
Owner 蔡剑前
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