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目的建立耐紫杉醇(PTX)子宫颈癌细胞株,观察N-乙酰半胱氨酸(NAC)对子宫颈癌细胞紫杉醇耐药的影响。方法用PTX处理并培养Hela细胞,建立耐PTX子宫颈癌细胞株Hela/PTX。观察和检测其细胞形态、细胞生长曲线、耐药指数、氧化还原状态及紫杉醇耐药基因1(Txr1)的表达。用NAC处理Hela/PTX细胞,观察NAC对其PTX耐药的影响。结果用PTX处理Hela细胞10个月后,可获得在500μg/L PTX中生长状态良好的Hela/PTX细胞。Hela/PTX细胞体积偏大,胞浆内颗粒较多,群体倍增时间是亲代细胞的1.32倍,对PTX具有显著耐药性,耐药指数为122.69。与Hela细胞相比,Hela/PTX细胞有高水平的活性氧(ROS)和Txr1 mRNA,但还原型谷胱甘肽(GSH)水平及超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶活性却有不同程度的降低,而氧化型谷胱甘肽(GSSG)水平和GSH/GSSG比值则无明显变化。NAC处理后,Hela/PTX细胞ROS和Txr1mRNA水平下降,对PTX敏感性增加。结论耐紫杉醇Hela/PTX细胞氧化还原状态失衡,Txr1表达增加。NAC能降低ROS水平,减少Txr1表达,可逆转其对PTX的耐受。
Objective To establish a paclitaxel-resistant (PTX) cervical cancer cell line and investigate the effect of N-acetylcysteine (NAC) on paclitaxel resistance in cervical cancer cells. Methods Hela cells were treated with PTX and cultured to establish PTX-resistant cervical cancer cell lines Hela / PTX. The cell morphology, cell growth curve, drug resistance index, redox status and the expression of paclitaxel resistance gene 1 (Txr1) were observed and detected. Hela / PTX cells were treated with NAC to observe the effect of NAC on PTX resistance. Results After Hela cells were treated with PTX for 10 months, Hela / PTX cells that grew well in 500 μg / L PTX were obtained. The volume of Hela / PTX cells was large, with more intracytoplasmic granules, doubling the population doubling time of the parental cells and having significant resistance to PTX, with a resistance index of 122.69. Hela / PTX cells had higher levels of reactive oxygen species (ROS) and Txr1 mRNA than Hela cells, but reduced glutathione (GSH) levels and superoxide dismutase, catalase and glutathione Peroxidase activity decreased to some extent, while the level of GSHG and GSH / GSSG did not change significantly. After NAC treatment, the levels of ROS and Txr1 mRNA in Hela / PTX cells decreased and the sensitivity to PTX increased. Conclusion The redox status of paclitaxel-resistant Hela / PTX cells is unbalanced and Txr1 expression is increased. NAC can reduce the level of ROS, reduce the expression of Txr1, can reverse its tolerance to PTX.