Effect of LHRH-PE40 on target cells via LHRH receptors.
OBJECTIVE
To detect the effect and cytotoxicity of luteinizing hormone-releasing hormone-Pseudomonas aeruginosa exotoxin 40 (LHRH-PE40) on target cells using LHRH receptors (LHRHR).


METHODS
The affinity of LHRH-PE40 and LHRH binding to LHRHR on the membrane surface of target cells were measured by enzyme linked immunosorbent assay. Morphological observations with light microscope were used to analyze its receptor pathway, with Spiegelmer, and cytotoxicity. IC(50) values of LHRH-PE40, which caused 50% inhibition of tumor cell growth were evaluated by MTT assay. The target cells were exposed to LHRH-PE40 and its cytotoxicity was analyzed by scanning and transmission electron microscopies, agarose gel electrophoresis, and flow cytometry.


RESULTS
LHRH-PE40 killed target cells by LHRHR pathway. The morphological changes in these cells showed decreased cell size, cytoplasmic membrane blebbing, and chromatin condensation and margination. At a certain concentration and time point, HeLa cells were also induced to undergo programmed cell death.


CONCLUSIONS
LHRH-PE40 induced target cells apoptosis via LHRHR.
Department of Biochemistry and Molecular Biology, China Medical University, Shenyang, People's Republic of China.
Xin
X
Deng
Sven
S
Klussmann
Guang-Mou
GM
Wu
Doreen
D
Akkerman
Ya-Qin
YQ
Zhu
Ying
Y
Liu
Hang
H
Chen
Ping
P
Zhu
Bing-Zhi
BZ
Yu
Guo-Li
GL
Zhang
D036002:ADP Ribose Transferases / Q000235:genetics*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001427:Bacterial Toxins / Q000235:genetics*; D001667:Binding, Competitive; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005098:Exotoxins / Q000235:genetics*; D005434:Flow Cytometry; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D006801:Humans; D008854:Microscopy, Electron; D011966:Receptors, LHRH / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D037521:Virulence Factors / Q000235:genetics*; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000970:Antineoplastic Agents; D001427:Bacterial Toxins; D005098:Exotoxins; D007987:Gonadotropin-Releasing Hormone; D011966:Receptors, LHRH; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A