The Establishment Of Vincristine-resistant Human Sarcoma Cell Line (ms5/vcr)

Objective: The aim of our study was to establish a vincristine(VCR)-resistant sarcoma cell line, which was derived from human embryonic muscle malignant transformed cells, named MS5, and observe their stem cell characteristics such as clonal ability and oncogenicity in vivo.Method:In this study, a cell line named MS5/VCR was established by gradually increasing concentration of VCR in vitro. The shape of the MS5 and MS5/VCR cells was observed in inverted microscope, the sensibility to VCR of the two cell lines were compared by MTT experiment. MS5 and MS5/VCR cells were injected i.p. into 8 nude mice, the mice were killed 55 day after injection and tumor formation was examined, immunohistochemical detection of mdr-1 was performed to observe its express in formation tumors.Result:After induction by VCR for about 7 months, MS5/VCR were obtained and they could proliferate steadily in medium with 6μg/ml vincristine. Compared with MS5, MS5/VCR were slightly round in shape, MTT experiment displayed that MS5/VCR cells was resistance to VCR. Clonal ability of MS5/VCR was stronger than that of MS5; Tumor size of MS5/VCR from nude mice were bigger than those of MS5, immunohistochemical study showed that mdr-1 could be detected in both MS5 and MS5/VCR tumors.Conclusions:1. MS5/VCR could be established by culture of MS5 with intermittently increasing of VCR concentration in the culture medium in vitro.2. Clone ability in vitro and oncogenicity in vivo of MS5/VCR resistant cells were stronger than that of MS5. Aim: The aim of the study was to identify the hemoglobin G-Chinese in 3 patients by hematological electrophoresis and DNA analysis.Methods: The presence of Hb variant was confirmed by cellulose acetate electrophoresis. DNA analysis based on the polymerase chain reaction (PCR), Pst I restriction enzyme analysis and sequencing was developed to confirm the presence of the mutation for the Hb G-Chinese.Results: 3 cases of Hb G-Chinese carrier were identified.Conclusions: The results showed that PCR was a simple, rapid, and inexpensive approach for the identification of Hb variant caused by gene mutation, the Gâ†'C substitution at Hb G-Chinese creates a Pst I restriction site.