Production and utilization of a novel anti-cancer drug in therapy
View Patent ↗This invention generally relates to a design and method for developing novel anti-tumor and/or anti-cancer drugs, vaccines and therapies, using microRNA and/or its shRNA homologs/mimics/derivatives. More specifically, the present invention relates to an use of a prokaryote-produced miRNA precursor (pro-miRNA) composition capable of being delivered into human cells and processed by the cells into mature miRNA effectors to elicit specific silencing effects on mir-302-targeted genes, subsequently leading to a beneficial result of tumor suppression and cancer therapy. The prokaryotic cells do not naturally express or process eukaryotic miRNA precursors (pre-miRNA); meanwhile, the present invention also teaches an inducible method for expressing pre-miRNAs, particularly mir-302 precursors by using the prokaryotic transcription system. Since mir-302 is a known tumor suppressor in human, this novel finding advances the design and method for developing new anti-cancer drugs, vaccines and/or therapies directed against multiple kinds of human tumors and cancers.
1. A method for inducing a gene silencing effect on BMI-1 and then leading to an increase of p16Ink4a or p14Arf tumor suppressor expression in vivo, comprising:
delivering a pro-mir-302 capable of being processed into at least one hairpin-like microRNA precursor in a human cell substrate to an amount, wherein said at least one hairpin-like microRNA precursor contains a seed sequence SEQ ID NO: 3 and said human cell substrate contains at least a cancerous cell type of human liver cancer, and wherein the amount of said hairpin-like microRNA precursor is higher than 200 micrograms per delivery in vivo.
2. The method of claim 1 , wherein said step of delivering said pro-mir-302 further comprises providing said pro-mir-302 from prokaryotic competent cells.
3. The method of claim 2 , wherein said pro-mir-302 is produced by said prokaryotic competent cells and is a precursor of mir-302.
4. The method of claim 2 , wherein said step of providing said pro-mir-302 further comprises isolating said pro-mir-302 from an extract or a lysate of said prokaryotic competent cells.
5. The method of claim 1 , wherein said step of delivering said pro-mir-302 capable of being processed into at least one hairpin-like microRNA precursor further comprises letting said pro-mir-302 be processed into mir-302a, mir-302b, mir-302c, and/or mir-302d in said human cell substrate.
6. The method of claim 1 , wherein said gene silencing effect further comprises an effect leading to reprogram high-grade malignant cancers to a relatively benign low-grade state.
7. The method of claim 1 , wherein said pro-mir-302 is delivered into said human cell substrate by a gene delivery method.
8. The method of claim 1 , wherein said pro-mir-302 is used as a drug for treating human liver cancer.
9. The method of claim 1 , wherein said pro-mir-302 is useful for pharmaceutical and/or therapeutic applications.