Application of genetically engineered bacterium VNP20009-M to preparation of medicine for treating sarcomas
A technology of VNP20009-M, genetically engineered bacteria, applied in gene therapy, microbial-based methods, anti-tumor drugs, etc., can solve the problem of no anti-tumor effect observed
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Embodiment 1
[0033] The construction of embodiment 1 genetically engineered bacteria VNP20009-M
[0034] 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.
[0035] (1) Construction of a plasmid expressing the L-methioninase gene.
[0036] 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:
[0037] 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 ...
Embodiment 2
[0048] Example 2 Effect of genetically engineered bacteria VNP20009-M in treating soft tissue sarcoma
[0049] 1) Past medical history and diagnosis
[0050] A male patient, 64 years old, had a mass in his left leg that was identified as spindle cell sarcoma by needle biopsy at Nanjing Drum Tower Hospital. After tumor resection, the tissue was taken again for pathological analysis, showing tumor cells SMA(-), DES(-), MyoD1(-), FN(+), STAT6(-), CD34(-), S100(-), CKpan (-), EMA (-), Vimentin (++), combined with HE section, consistent with malignant fibrous histiocytoma.
[0051] Periodic follow-up and re-examination of CT revealed a soft tissue mass in the upper lobe of the right lung, and biopsy pathology showed spindle cell sarcoma. Based on the patient's previous treatment and related examinations, a diagnosis of postoperative recurrence of spindle cell sarcoma with lung metastasis was made.
[0052] 2) Treatment plan
[0053] Genetically engineered bacteria VNP20009-M di...
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