PMID- 26718643
OWN - NLM
STAT- MEDLINE
DCOM- 20161017
LR  - 20220718
IS  - 1791-2431 (Electronic)
IS  - 1021-335X (Linking)
VI  - 35
IP  - 2
DP  - 2016 Feb
TI  - Expression and purification of recombinant ATF-mellitin, a new type fusion 
      protein targeting ovarian cancer cells, in P. pastoris.
PG  - 1179-85
LID - 10.3892/or.2015.4448 [doi]
AB  - Melittin is well known to possess cytolytic activity with wide-spectrum lytic 
      properties and its potential use as an agent to treat several types of cancer has 
      been tested. Due to the non-specific toxicity, melittin can impair not only 
      cancer cells but also normal tissue. Thus, tumor-targeted toxins may be helpful 
      for developing novel anticancer therapeutics. The urokinase-type plasminogen 
      activator (uPA) plays a central role in tissue remodelling events occurring in 
      normal physiology and in pathophysiology, including cancer invasion and 
      metastasis. Heartening findings showed that uPA receptor is predominantly 
      expressed on many types of cancer. Therefore, the amino-terminal fragment (ATF) 
      of uPA which was able to identify and bond with cancer cells was used as the 
      cell-targeting domain to make up tumor-targeted toxin in this study. In the 
      present study, pPICZalphaC-ATF-melittin eukaryotic expression vector was successfully 
      constructed. After transformed into P. pastoris and induced by methanol, 
      rATF-mellitin was detected by SDS-PAGE and western blot analysis. After induction 
      with methanol, the expression level of rATF-mellitin was 312 mg/l in 80-l 
      fermentor. rATF‑mellitin was purified to >95% purity using SP Sepharose ion 
      exchange chromatography and source 30 RPC with 67.2% recovery. Cell 
      proliferation assay showed that rATF-melittin inhibited growth of SKOV3 cells and 
      had no cytotoxicity effect on normal cells. For the first time, we established a 
      stable and effective rATF-mellitin P. pastoris expression system to obtain a high 
      level of expression of secreted rATF-mellitin which was purified by a highly 
      efficient purification procedure.
FAU - Su, Manman
AU  - Su M
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
FAU - Chang, Weiqin
AU  - Chang W
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
FAU - Zhang, Kun
AU  - Zhang K
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
FAU - Cui, Manhua
AU  - Cui M
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
FAU - Wu, Shuying
AU  - Wu S
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
FAU - Xu, Tianmin
AU  - Xu T
AD  - Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, 
      Changchun, Jilin 130041, P.R. China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151125
PL  - Greece
TA  - Oncol Rep
JT  - Oncology reports
JID - 9422756
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Naphthoquinones)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Receptors, Urokinase Plasminogen Activator)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 6N4FA2QQ6A (beta-lapachone)
RN  - Y4S76JWI15 (Methanol)
SB  - IM
MH  - Antineoplastic Agents/*isolation & purification/metabolism/pharmacology
MH  - Cell Line, Tumor
MH  - Drug Screening Assays, Antitumor
MH  - Female
MH  - Gene Expression/drug effects
MH  - Genes, Synthetic
MH  - Genetic Vectors/*genetics
MH  - Humans
MH  - Methanol/pharmacology
MH  - Molecular Targeted Therapy
MH  - Naphthoquinones/*isolation & purification/metabolism/pharmacology
MH  - Neoplasm Proteins/metabolism
MH  - Ovarian Neoplasms/*pathology
MH  - Pichia/*metabolism
MH  - Plasmids/*genetics
MH  - Receptors, Urokinase Plasminogen Activator/metabolism
MH  - Recombinant Fusion Proteins/biosynthesis/isolation & purification/pharmacology
MH  - Transformation, Genetic
EDAT- 2016/01/01 06:00
MHDA- 2016/10/19 06:00
CRDT- 2016/01/01 06:00
PHST- 2015/08/04 00:00 [received]
PHST- 2015/09/22 00:00 [accepted]
PHST- 2016/01/01 06:00 [entrez]
PHST- 2016/01/01 06:00 [pubmed]
PHST- 2016/10/19 06:00 [medline]
AID - 10.3892/or.2015.4448 [doi]
PST - ppublish
SO  - Oncol Rep. 2016 Feb;35(2):1179-85. doi: 10.3892/or.2015.4448. Epub 2015 Nov 25.