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Production and cytotoxicity against cancer cells of recombinant cecropin b-polyproline |
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| รหัสดีโอไอ | |
| Title | Production and cytotoxicity against cancer cells of recombinant cecropin b-polyproline |
| Creator | Sirin Sarunyutanon |
| Contributor | Sarintip Sooksai, Kittinan Komolpis |
| Publisher | Chulalongkorn University |
| Publication Year | 2558 |
| Keyword | Peptide antibiotics, Recombinant proteins, เปปไทด์ต้านจุลชีพ, รีคอมบิแนนต์โปรตีน |
| Abstract | Cecropin B (CB) is a natural anticancer antimicrobial peptide which has the highest antibacterial activity among the Cecropin family. In this work, Cecropin B - polyproline (CB-PP) was constructed by adding polyproline (PP) which is a part of progesterone receptor (PR) to the C-terminus of CB sequence. The CB and CB-PP gene fragments were cloned into the pPICZαA expression vector. Then, the recombinant plasmid CB and CB-PP were transformed into Pichia pastoris strain KM71H by electroporation. The recombinant protein CB (rCB) and CB-PP (rCB-PP) were successfully produced by inducing with 0.5% methanol for 24 and 48 hours and purified using nickel column chromatography. The size of rCB and rCB-PP were found to be approximately 4.9 and 6.6 kDa, respectively. The cytotoxicity against six cell lines including human breast carcinoma (BT-474), human lung bronchus carcinoma (ChaGo-K1), human liver hepatocellular carcinoma (Hep-G2), human gastric carcinoma (Kato-III), human colon carcinoma (SW-620) and human lung fibroblast (WI-38) were investigated. All cell lines were treated with recombinant protein CB and CB-PP with the protein concentration between 0.078 µM – 20 µM for 72 hours. The results revealed that more than 50% of those five cancer cells were dead at the rage of 7 µM - 20 µM protein concentrations while more than 80% of normal cells was alive even treated with the same protein concentration. Those resultsmindicated that rCB และ rCB-PP could be produced by the transformed Pichia pastoris and the obtained recombinant proteins exhibited anticancer activity. |
| URL Website | cuir.car.chula.ac.th |
