Application of genetically engineered bacterium VNP20009-M to preparation of lung cancer prevention and treatment drugs
A VNP20009-M, a technology of genetically engineered bacteria, applied in gene therapy, microorganism-based methods, drug combinations, etc., can solve problems such as no observed anti-tumor effect
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Embodiment 1
[0039] The construction of embodiment 1 genetically engineered bacteria VNP20009-M
[0040] The construction method and process of the genetically engineered bacteria VNP20009-M of the present invention are as described in the examples of Chinese patent CN105983103A.
[0041] (1) Construction of a plasmid expressing the L-methioninase gene.
[0042]Synthesized L-methioninase (GenBank: L43133.1) gene was subcloned into pUC57 plasmid (GenScript), and then subcloned into pSVSPORT plasmid (invitrogen) through Kpn I and Hind III restriction sites to obtain pSVSPORT-L- methioninase expression plasmid. The specific construction process is as follows:
[0043] Digest the pSVSPORT plasmid with Kpn I and Hind III. The digestion system is: 2 μg plasmid DNA, 3 μL 10×buffer, 1.5 μL Kpn I enzyme, 1.5 μL Hind III enzyme, add ddH2O to make up the volume to 30 μL, and incubate at 37°C for 3 hours . Then the digestion system was separated by electrophoresis in 1% agarose gel, the DNA band w...
Embodiment 2
[0054] The antitumor effect of embodiment 2 genetically engineered bacteria VNP20009-M
[0055] 1), past medical history and diagnosis
[0056] A male patient, 73 years old, underwent thoracoscopic radical resection of squamous cell carcinoma of the lower lobe of the right lung 5 months after a new mass was found in the neck (such as Figure 4 As shown), the size of the lesion at the neck was measured to be about 8cm×9cm. Biopsy of the mass site identified as cancerous by cell smear (eg, Figure 5 shown), and skeletal ECT showed active bone metabolism in multiple places, and chest CT examination reported sternal destruction with mass formation.
[0057] According to the patient's previous treatment and related examinations, the patient was diagnosed with recurrence and metastasis of right lower lobe squamous cell carcinoma after surgery, but there is no standard clinical treatment plan.
[0058] 2) Treatment plan
[0059] The diluted VNP20009-M was evenly injected into the...
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