ABSTRACT
In our study, a water-soluble polysaccharide (CCPa-1) was successfully purified from the fruiting bodies of Coprinus comatus by DEAE-cellulose and Sepharose CL-6B column chromatography. The molecular weight was evaluated to be 53.6 kDa as determined by high performance size exclusion chromatography (HPSEC). Sugar composition analysis revealed that CCPa-1 consisted primarily of galactose, glucose and arabinose in a molar ratio of 6.6:1.2:2.2. CCPa-1 could not only inhibit the growth of sarcoma 180 (S180) tumor transplanted in mice, but also increase the relative spleen/thymus indexes and body weight of tumor bearing mice. Moreover, Con A- or LPS-induced splenocyte proliferation was also enhanced after CCPa-1 administration in tumor-bearing mice. Furthermore, CCPa-1 significantly enhanced the Con A- or LPS-induced splenocyte proliferation and increased the production of TNF-α and IL-2. All the data demonstrated that CCPa-1 had a potential application as natural antitumor agent with immunomodulatory activity.
Subject(s)
Antineoplastic Agents/pharmacology , Coprinus/chemistry , Fungal Polysaccharides/pharmacology , Immunologic Factors/pharmacology , Animals , Antineoplastic Agents/isolation & purification , Cell Proliferation/drug effects , Fungal Polysaccharides/isolation & purification , Immunologic Factors/isolation & purification , Interleukin-2/blood , Male , Mice , Mice, Inbred ICR , Sarcoma, Experimental/drug therapy , Sarcoma, Experimental/immunology , Sarcoma, Experimental/pathology , Spleen/drug effects , Spleen/pathology , Tumor Necrosis Factor-alpha/bloodABSTRACT
AIM: To construct the eukaryotic recombinant expression plasmid of pcDNA3.1(+)-PRMT1. METHODS: Human PRMT1 cDNA was amplified by reverse transcription polymerase chain reaction (RT-PCR). After digested by BamH I, Hind III and ligation, PRMT1 was inserted into pcDNA3.1(+)eukaryotic expression vector. The positive colonies were screened and identified by PCR and sequencing. pcDNA3.1(+)-PRMT1 plasmid was then transfected into the cultured A549 cell line with Lipofectamine(TM);2000. Realtime-PCR and Western blot were used to detect the mRNA and protein expression of PRMT1 respectively. RESULTS: The PRMT1 cDNA was successfully amplified, and pcDNA3.1(+)-PRMT1 were constructed. The inserted sequence in pcDNA3.1(+)-PRMT1 was the same as the sequence of PRMT1 cDNA published in NCBI GenBank. Further, Realtime PCR and Western blot results validated the recombinant plasmid expressed in A549 cell line efficiently. CONCLUSION: pcDNA3.1(+)-PRMT1 recombinant was successfully constructed.
