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Application of cisplatin combined with hTERT-HRP/IAA in medicament for treating cervical cancer
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A cervical cancer, cisplatin technology, applied in the field of medicine, can solve the problems of decreased combined killing effect, decreased telomerase activity, etc.
Inactive Publication Date: 2010-11-17
WUHAN UNIV
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It was found that the combined use of cisplatin or carmustine at the half inhibitory rate (IC50) of them can reduce the activity of telomerase in the cells, resulting in a decrease in the combined killing effect
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[0034] 1.1 Cells: Human cervical cancer cells (Hela, CCTCC number GDC009) were purchased from the China Center for Type Culture Collection (CCTCC), and routinely cultured and passaged.
[0035] 1.2 Plasmid: The plasmid phTERTp-Luc has been successfully constructed by the applicant (see references: Liao ZK, Zhou FX, Luo ZG, Zhang WJ, Xiong J, Bao J, Han G, Zhang MS, Xie CH, Zhou YF. Radio- Activation of hTERT promoter in larynxsquamous carcinoma cells: An'indirected-activator'strategyin radio-gene-therapy.Oncol Rep.19(2008)281-286.), internal reference plasmid pRL-TK and control plasmid pGL3-Basic were purchased from PROMEGA company .
[0036] 1.3 Reagents: Liposome METAFECTENE was purchased from Biontex Company, and the dual fluorescence detection kit was purchased from PROMEGA Company. DDP was purchased from Dezhou Deyao Pharmaceutical Co., Ltd., and RPMI-1640 cu...
Embodiment 2
[0044] Example 2 The killing effect of cisplatin chemotherapy combined with phTERTp-HRP / IAA on cervical cancer cells
[0045] 1 Experimental materials:
[0046] 1.1 Cells: Cells: Human cervical cancer cells (Hela, CCTCC number GDC009) were purchased from the China Center for Type Culture Collection (CCTCC), and routinely cultured and passaged.
[0047] 1.2 Plasmid: Plasmid phTERTp-HRP has been successfully constructed by the applicant (see references: Liao ZK, Zhou FX, Luo ZG, Zhang WJ, Xiong J, Bao J, Han G, Zhang MS, Xie CH, Zhou YF. Radio- Activation of hTERT promoter in larynxsquamous carcinoma cells: An'indirected-activator'strategy in radio-gene-therapy. Oncol Rep.19(2008)281-286).
[0048] 1.3 Reagents: Liposome METAFECTENE was purchased from Biontex Company, and the dual fluorescence detection kit was purchased from PROMEGA Company. Indole acetic acid was purchased from Fluka, the apoptosis kit AnnexinV-FITC kit was purchased from Jingmei Bioengineering Co., Ltd., R...
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Abstract
The invention discloses an application of cisplatin combined with phTERTp-HRP / IAA in drugs for the treatment of cervical cancer cells, in particular to the strengthening role of the cisplatin on the activity of a hTERT genepromoter. The cisplatin and the phTERTp-HRP / IAA are commonly used in the synergy for killing the human cervical cancer cells, and the action mechanism thereof is further explored. The application adopts Hela cells for carrying out the study and evaluates the impacts of the cisplatin on the activity of the hTERT promoter through the expression of downstream luciferasereporter gene regulated by the hTERT promoter. The synergic killing role and the mechanism of the cisplatin and the hTERT promoter during the combined use for regulating the gene therapy of an HRP / IAA system are evaluated through the detection of cellproliferation rate, cellapoptosis rate and cell cycle. The results confirm that the cisplatin can strengthen the activity of the hTERT gene promoter; and when in combined use with the phTERTp-HRP / IAA, the killing role of the human cervical cancer cells is significant.
Description
technical field [0001] The present invention relates to the technical field of medicine, and more specifically relates to the application of cisplatin in the medicine for enhancing the gene therapy mediated by the telomerasereverse transcriptasegene (hTERT) promoter, and cisplatin can enhance the activity of the hTERT promoter and enhance the expression of downstream genes role in. At the same time, it also relates to the application of cisplatin and hTERT promoter to regulate horseradish peroxidase (HRP) / indole acetic acid (IAA) system to produce synergistic effect to destroy cervical cancer cells and inhibit the growth of cervical cancer cells. Background technique [0002] Looking at the changing pattern of the disease spectrum at home and abroad, malignant tumors have become a major disease that seriously threatens human health. With the advancement of molecular biology, gene therapy has brought hope to defeat tumors. Various countries have carried out a lot of resea...
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