[Induction of apoptosis in various cancer cell lines by human soluble TRAIL expressed in Escherichia coli].
OBJECTIVE
To investigate the induction of apoptosis by human soluble TRAIL expressed in Escherichia coli and analyze its relationship with p53 in various cancer cell lines, especially the solid tumor.


METHODS
TRAIL codons 114 approximately 281 were amplified by polymerase chain reaction. The soluble segment was expressed in Escherichia coli JF1125, refolded and purified. Induction of apoptosis was assessed by examination of morphological changes under electron microscope and quantified by FACS analysis. The mutation of p53 in employed cell lines was detected by immunohistochemistry.


RESULTS
By multi-step purification, the purity of human soluble TRAIL exceeded 90%. Induction of apoptosis by the product exhibited a marked increase in human breast cancer cell line MCF-7, HeLa cervical carcinoma cells and hepatocellular carcinoma cell lines SMMC7721, BEL7402, and BEL7405. The mutation of p53 in employed cell lines was positive in aforementioned cell lines.


CONCLUSIONS
The human soluble TRAIL expressed in Escherichia coli has similar activity to that generated in cells to induce apoptosis in various cancer cell lines, and acts independently of p53.
Zheng
X
X
Liver Cancer Institute, Shanghai Medical University, Shanghai 200032, China.
Guan
X
X
Lin
Z
Z
D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D004926:Escherichia coli; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D012995:Solubility; D053221:TNF-Related Apoptosis-Inducing Ligand; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
D051017:Apoptosis Regulatory Proteins; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; D016159:Tumor Suppressor Protein p53