AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horsesh...AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horseshoe crab(Tachypleustridentatus)hemocytes.BGC-823 cells and thecells treated with 2.0mg/L tachyplesin wereexamined respectively under light microscope,scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone therestorational alteration in morphology andultrastructure after tachyplesin treatment.Thechanges were as follows:the shape of cells wasunanimous,the volume enlarged and cellsturned to be flat and spread,the nucleo-cytoplasmic ratio lessened and nuclear shapebecame rather regular,the number of nucleolusreduced and its volume lessened,heter-chromatin decreased while euchromatinincreased in nucleus.In the cytoplasm,mitochondria grew in number with consistentstructure relatively,Golgi complex turned to betypical and well-developed,rough endoplasmicreticulum increased and polyribosomedecreased.The microvilli at cellular surfacewere rare and the filopodia reduced whilelamellipodia increased at the cell edge.CONCLUSION Tachyplesin could alter themalignant morphological and ultrastructuralcharacteristics of human gastric carcinoma cellseffectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.展开更多
AIM: To investigate the effects of tachyplesin on the cell cycle regulation in human hepatcarcinoma cells.METHODS: Effects of tachyplesin on the cell cycle in human hepatocarcinoma SMMC-7721 cells were assayed with fl...AIM: To investigate the effects of tachyplesin on the cell cycle regulation in human hepatcarcinoma cells.METHODS: Effects of tachyplesin on the cell cycle in human hepatocarcinoma SMMC-7721 cells were assayed with flow cytometry. The protein levels of p53, p16, cyclin D1 and CDK4 were assayed by immunocytochemistry. The mRNA levels of p21WAF1/CIP1 and c-myc genes were examined with in situ hybridization assay.RESULTS: After tachyplesin treatment, the cell cycle arrested at G0/G1 phase, the protein levels of mutant p53, cyclin D1 and CDK4 and the mRNA level of c-myc gene were decreased, whereas the levels of p16 protein and p21wWF1/CIP1 mRNA increased.CONCLUSION: Tachyplesin might arrest the cell at G0/G1 phase by upregulating the levels of p16 protein and p21WAF1/CIP1 mRNA and downregulating the levels of mutant p53, cyclin D1 and CDK4 proteins and c-myc mRNA, and induce the differentiation of human hepatocacinoma cells.展开更多
AIM: To investigate the antitumor activities of tachyplesin on human hepatocellula r carcinoma (HCC) cells.METHODS: Tachyplesin, isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes...AIM: To investigate the antitumor activities of tachyplesin on human hepatocellula r carcinoma (HCC) cells.METHODS: Tachyplesin, isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes, was used to treat the human HCC cell line SMMC-7721. Effects of tachyplesin on the proliferation of SMMC-7721 cells were measured with trypan blue dye exclusion test and HE staining. The morphology and ultrastructure of the cells were examined by light microscopy and transmission electron microscopy, respectively. The activities of γ-glutamyltransferase (γ-GT) and tyrosine aminotransferase (TAT) were assayed with biochemical methods. The levels of alpha fetoprotein (α-FP), proliferating cell nuclear antigen (PCNA), p21wAF1/CIP1 and c-myc were examined by immunocytochemistry. RESULTS: After treatment with tachyplesin 3.0 mg/L, the proliferation of SMMC-7721 cells was inhibited significantly, with the cell growth inhibitory rate amounted to 55.57 % and the maximum cell mitotic index declined by 43.68 %. The morphology and ultrastructure underwent restorational alteration. The activity of γ-GT declined while TAT activity increased obviously, and the levels of α-FP and PCNA decreased. Moreover, the expression of p21WAF1/CIP1 protein was upregulated and that of c-myc protein was down-regulated. CONCLUSION: Tachyplesin could effectively inhibit the proliferation of hepatocarcinoma cells, reverse the malignant morphological and ultrastructural characteristics, alter the levels of enzymes and antigens, regulate the expression of differentiation-associated oncogene and tumor suppressor gene, and induce hepatocarcinama cell differentiation.展开更多
This study aimed to investigate the synergism of the TAT PTD （ protein transduction domain in HIV-1 transactivator of transcription protein） to antibacte- rial peptide tachyplesin from Tachp/eus tr/dentatus. Treated...This study aimed to investigate the synergism of the TAT PTD （ protein transduction domain in HIV-1 transactivator of transcription protein） to antibacte- rial peptide tachyplesin from Tachp/eus tr/dentatus. Treated with Pichia pastoris preferred codon optimization, using the eDNA sequence of tuchyplesin mature pep- tide （54 aa） harboring TAT FFD sequence （11 aa） as reference template, six single-stranded oligonueleotides were designed, the sequences of restriction sites EcoR I and Xba I were introduced to the 5＇ end of primers P1 and P6, respectively. TAT PTD ＋ Tachyplesin fusion gene with a full length of 219 bp was artificially synthesized by overlap extension PCR, which laid the preliminary foundation for subsequent functional and synergism studies.展开更多
Tachyplesin gene was designed and chemically synthesized with codon usage bias. The stop codon TAG and some restriction sites convenient for further cloning were added to this gene. The synthesized gene cloned into ye...Tachyplesin gene was designed and chemically synthesized with codon usage bias. The stop codon TAG and some restriction sites convenient for further cloning were added to this gene. The synthesized gene cloned into yeast expression vector pPIC9 (with α_secretion signal) was transformed into host strain GS115 by electroporation. The tachyplesin expressed from recombinants Y PIC27 and Y PIC42 showed an inhibition activity against the germination of the spores of \%Magnaporthe grisea\%. Southern blot performed with chromosome genome of the two recombinants indicated a single copy of the expression cassette was integrated at the chromosomal AOX 1 locus by which the genomic AOX 1 gene was functionally disrupted and Northern blot showed the presence of transcripts of the tachyplesin gene.展开更多
基金the Natural Science Foundation of Fujian Province,No.C97015
文摘AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horseshoe crab(Tachypleustridentatus)hemocytes.BGC-823 cells and thecells treated with 2.0mg/L tachyplesin wereexamined respectively under light microscope,scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone therestorational alteration in morphology andultrastructure after tachyplesin treatment.Thechanges were as follows:the shape of cells wasunanimous,the volume enlarged and cellsturned to be flat and spread,the nucleo-cytoplasmic ratio lessened and nuclear shapebecame rather regular,the number of nucleolusreduced and its volume lessened,heter-chromatin decreased while euchromatinincreased in nucleus.In the cytoplasm,mitochondria grew in number with consistentstructure relatively,Golgi complex turned to betypical and well-developed,rough endoplasmicreticulum increased and polyribosomedecreased.The microvilli at cellular surfacewere rare and the filopodia reduced whilelamellipodia increased at the cell edge.CONCLUSION Tachyplesin could alter themalignant morphological and ultrastructuralcharacteristics of human gastric carcinoma cellseffectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.
基金the National Natural Science Foundation of China,No.30170724
文摘AIM: To investigate the effects of tachyplesin on the cell cycle regulation in human hepatcarcinoma cells.METHODS: Effects of tachyplesin on the cell cycle in human hepatocarcinoma SMMC-7721 cells were assayed with flow cytometry. The protein levels of p53, p16, cyclin D1 and CDK4 were assayed by immunocytochemistry. The mRNA levels of p21WAF1/CIP1 and c-myc genes were examined with in situ hybridization assay.RESULTS: After tachyplesin treatment, the cell cycle arrested at G0/G1 phase, the protein levels of mutant p53, cyclin D1 and CDK4 and the mRNA level of c-myc gene were decreased, whereas the levels of p16 protein and p21wWF1/CIP1 mRNA increased.CONCLUSION: Tachyplesin might arrest the cell at G0/G1 phase by upregulating the levels of p16 protein and p21WAF1/CIP1 mRNA and downregulating the levels of mutant p53, cyclin D1 and CDK4 proteins and c-myc mRNA, and induce the differentiation of human hepatocacinoma cells.
基金the National Natural Science Foundation of China,No.30170724Natural Science Foundation of Fujian Province,No.C97015
文摘AIM: To investigate the antitumor activities of tachyplesin on human hepatocellula r carcinoma (HCC) cells.METHODS: Tachyplesin, isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes, was used to treat the human HCC cell line SMMC-7721. Effects of tachyplesin on the proliferation of SMMC-7721 cells were measured with trypan blue dye exclusion test and HE staining. The morphology and ultrastructure of the cells were examined by light microscopy and transmission electron microscopy, respectively. The activities of γ-glutamyltransferase (γ-GT) and tyrosine aminotransferase (TAT) were assayed with biochemical methods. The levels of alpha fetoprotein (α-FP), proliferating cell nuclear antigen (PCNA), p21wAF1/CIP1 and c-myc were examined by immunocytochemistry. RESULTS: After treatment with tachyplesin 3.0 mg/L, the proliferation of SMMC-7721 cells was inhibited significantly, with the cell growth inhibitory rate amounted to 55.57 % and the maximum cell mitotic index declined by 43.68 %. The morphology and ultrastructure underwent restorational alteration. The activity of γ-GT declined while TAT activity increased obviously, and the levels of α-FP and PCNA decreased. Moreover, the expression of p21WAF1/CIP1 protein was upregulated and that of c-myc protein was down-regulated. CONCLUSION: Tachyplesin could effectively inhibit the proliferation of hepatocarcinoma cells, reverse the malignant morphological and ultrastructural characteristics, alter the levels of enzymes and antigens, regulate the expression of differentiation-associated oncogene and tumor suppressor gene, and induce hepatocarcinama cell differentiation.
基金Supported by Natural Science Foundation of Guangdong Province(06025389)Post-doctoral Project of Fujian Province
文摘This study aimed to investigate the synergism of the TAT PTD （ protein transduction domain in HIV-1 transactivator of transcription protein） to antibacte- rial peptide tachyplesin from Tachp/eus tr/dentatus. Treated with Pichia pastoris preferred codon optimization, using the eDNA sequence of tuchyplesin mature pep- tide （54 aa） harboring TAT FFD sequence （11 aa） as reference template, six single-stranded oligonueleotides were designed, the sequences of restriction sites EcoR I and Xba I were introduced to the 5＇ end of primers P1 and P6, respectively. TAT PTD ＋ Tachyplesin fusion gene with a full length of 219 bp was artificially synthesized by overlap extension PCR, which laid the preliminary foundation for subsequent functional and synergism studies.
文摘Tachyplesin gene was designed and chemically synthesized with codon usage bias. The stop codon TAG and some restriction sites convenient for further cloning were added to this gene. The synthesized gene cloned into yeast expression vector pPIC9 (with α_secretion signal) was transformed into host strain GS115 by electroporation. The tachyplesin expressed from recombinants Y PIC27 and Y PIC42 showed an inhibition activity against the germination of the spores of \%Magnaporthe grisea\%. Southern blot performed with chromosome genome of the two recombinants indicated a single copy of the expression cassette was integrated at the chromosomal AOX 1 locus by which the genomic AOX 1 gene was functionally disrupted and Northern blot showed the presence of transcripts of the tachyplesin gene.
