Gene medicine for promoting differentiation of tumor stem cells and applications thereof

A technology of tumor stem cells and gene medicine, applied in the field of gene medicine that promotes the differentiation of tumor stem cells, can solve the problems of recurrence and metastasis that plague surgeons, produce tumor tissue treatment resistance, and lack of differentiation of tumor stem cells, so as to improve drug sensitivity, Safe to use, low toxicity and side effects

Inactive Publication Date: 2013-09-11

SHANGHAI PULMONARY HOSPITAL

1 Cites 0 Cited by

AI-Extracted Technical Summary

Problems solved by technology

[0002] Traditional tumor treatment methods include chemotherapy, radiotherapy and surgery, but from the perspective of clinical treatment effects, none of them can completely cure the tumor

Moreover, due to the non-specific killing effect of radiotherapy and chemotherapy, the toxicity to the body is relatively high, and long-term use will cause treatment tolerance of tumor tissues.

The curative effect of surgery...

Abstract

The invention provides a gene medicine for promoting the differentiation of tumor stem cells. The gene medicine comprises a recombinant of which the target gene is a Nell-1 gene and can effectively induce the differentiation of the tumor stem cells, inhibit the multiplication of tumor cells, reduce the drug resistance of the tumor cells and improve the drug sensitivity and can be combined with chemical treatment to enhance the chemical treatment effect; and the gene medicine is low in toxic and side effects and safely used. A carrier of the recombinant can be an expression plasmid or a virus carrier, preferably the virus carrier; the virus carrier is preferably selected from one of an adenovirus carrier, a herpes simplex virus carrier, a retroviruse carrier, an adeno associated virus carrier and a slow virus carrier; and most preferably, the virus carrier is the adenovirus carrier.

Application Domain

Genetic material ingredientsAntineoplastic agents +1

Technology Topic

Adeno associate virusSide effect +12

Image

Examples

Experimental program(4)

Example Embodiment

[0024] Example 1 Nell-1 gene applied to glioma

[0025] The Nell-1 gene sequence was obtained by RT-PCR. After the detection was correct, the full-length gene was amplified by PCR in vitro, and then the amplified product was cloned into an expression plasmid to obtain an expression plasmid (Nell-1) carrying the Nell-1 gene. 1 expression plasmid). The Nell-1 expression plasmid was used to transfect into glioma stem cell line (DA66), and the positive ratio of CD133 in this tumor stem cell line was about 65-70%.

[0026] After G418 screening, a stable and high Nell-1 expression cell line (DA66-Nell-1) was obtained, stained with CD133 antibody, and then analyzed by flow cytometry, such as figure 1 As shown (where DA66 is a complete blank control without any treatment, and DA66-Neo is a blank plasmid transfection), Nell-1 overexpression significantly reduces the expression of CD133, while the transfection of blank plasmid (DA66-Neo) significantly reduces the expression of CD133. The expression of CD133 had little effect, indicating that overexpression of Nell-1 suppressed the expression of CD133.

[0027] DA66-Nell-1 and DA66-Neo were cultured in vitro, and their differentiation was observed. The results are as follows figure 2 As shown, glioma stem cells overexpressed Nell-1, leading to the differentiation of tumor stem cells. Markers related to neural stem cell differentiation (GFAP, MAP2) and bone differentiation (OPN and OCN) were also significantly increased in DA66-Nell-1 ( figure 2 A), at the same time, the proliferation ability of DA66-Nell-1 cells was significantly reduced in vitro ( figure 2 B), indicating that Nell-1 can effectively induce glioma stem cell differentiation and inhibit cancer cell proliferation.

[0028] Male nude mice aged about 6-8 weeks were selected and divided into two groups. DA66-Neo or DA66-Nell-1 cells were subcutaneously injected respectively. After 4 weeks, the tumor tissue was taken and the tissue was stained with HE. The results are as follows: image 3 As shown, the tumorigenic ability of DA66-Nell-1 was significantly less than that of the DA66-Neo control group ( image 3 A). HE staining revealed that tumors formed after injection of Nell-1-overexpressing cells were characterized by differentiation and mineralization ( image 3 B).

[0029] In vitro drug sensitivity assay of DA66-Nell-1 and DA66-Neo cells, the results are as follows Figure 4 As shown, after Nell-1 overexpression in DA66-Nell-1, tumor cells lost their resistance to chemotherapeutic drugs. It can be seen that Nell-1 can not only induce the differentiation of tumor stem cells, but also enhance the sensitivity of tumor stem cells to chemotherapy. Spend.

[0030] Cancers other than gliomas are tested the same way as above, and the test results are similar to gliomas.

Example Embodiment

[0031] The preparation of the recombinant of embodiment 2 target gene is Nell-1

[0032] The Nell-1 gene sequence was obtained by RT-PCR. After the detection was correct, the full length of the gene was amplified by PCR in vitro, and the amplified product clone and adenovirus shuttle plasmid were cut with specific endonuclease to construct a Nell-carrying plasmid. -1 gene shuttle plasmid, then subcloned into pQB1-AdCMV5 (containing adenovirus E1A), co-transfected with QB1-viral DNA, positive virus clones were screened by PCR to obtain Ad-Nell-1 recombinant, amplified, transfected HEK293 cells were transfected, cultured, the Nell-1 gene recombinant adenovirus was roughly divided, purified by ultracentrifugation, sterilized, and obtained.

Example Embodiment

[0033] The preparation of embodiment 3 injection

[0034] The Ad-Nell-1 recombinant obtained in Example 2 was 2×10 15 Virus titer, add normal saline for injection to 1000mL, sterile filter, and dispense into 2mL vials.

PUM

Description & Claims & Application Information

We can also present the details of the Description, Claims and Application information to help users get a comprehensive understanding of the technical details of the patent, such as background art, summary of invention, brief description of drawings, description of embodiments, and other original content. On the other hand, users can also determine the specific scope of protection of the technology through the list of claims; as well as understand the changes in the life cycle of the technology with the presentation of the patent timeline. Login to view more.

Pharmaceutical composition for preventing or treating cancer, containing cd300c expression inhibitor or activity inhibitor

PendingUS20210238596A1inhibit proliferationincrease in number

Owner:CENTRICSBIO INC

Application of polyethylene glycol artesunate in preparation of antitumor drug

InactiveCN106727499AThe dose-effect relationship is obviousInhibit proliferation

Owner:KPC PHARM INC

Classification and recommendation of technical efficacy words

Inhibit proliferation
reduce drug resistance

Gene medicine for promoting differentiation of tumor stem cells and applications thereof

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Image

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

Similar technology patents

Indole derivative as well as synthesis method and application thereof

Colorectal cancer targeted therapy drug

Antibodies for diagnosis and therapeutic treatment of prostate cancer

Pharmaceutical composition for preventing or treating cancer, containing cd300c expression inhibitor or activity inhibitor

Application of polyethylene glycol artesunate in preparation of antitumor drug

Classification and recommendation of technical efficacy words

Humanized anti-EGF receptor monoclonal antibody

Novel physiologically active substance

Single-stranded circular RNA and DNA and preparation method and application thereof

Fluorinated Arylamide Derivatives

Application of dihydromyricetin (DHM) in preparation of anti-hepatoma medicines

Fungicidal composition containing prochloraz manganese complex

Oral liquid for treating air sacculitis caused by colibacillosis of chicken and preparation method thereof

Pharmaceutical composition for treating heart failure with preserved ejection fraction and application of pharmaceutical composition

Method of non-chemical treatment for preventing and controlling insects of grain storage

Pesticide preparation for reducing drug resistance of plant diseases and insect pests