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目的:观察在阿霉素诱导大肠癌LoVo细胞产生多药耐药过程中,端粒酶逆转录酶基因表达,明确端粒酶是否参与了多药耐药机制.方法:采用阿霉素浓度递增法,建立人大肠癌细胞多药耐药模型LoVo/Adr;用MTT法鉴定耐药LoVo/Adr细胞的耐药性;以流式细胞术检测其周期分布;用RT-PCR方法检测hTERT mRNA水平在诱导耐药过程中的变化.结果:历经8 mo+、传代67次连续培养,建立了人大肠癌耐药模型LoVo/Adr细胞株;LoVo/Adr对阿霉素、长春新碱、丝裂霉素、环磷酰胺和5-氟尿嘧啶耐药倍数分别是61倍、14倍、3倍、9倍和1倍;LoVo/Adr细胞与LoVo细胞相比,S期细胞减少,而G1,G2期增多;亲本LoVo细胞的hTERT mRNA呈低水平表达,在阿霉素诱导初期即显著增高,随后基本保持高表达状态,并不随着耐药性增加而增强.结论:耐药株LoVo/Adr是一个典型的具有多药耐药性表型的耐药模型,端粒酶参与了其耐药机制.
OBJECTIVE: To observe the expression of telomerase reverse transcriptase gene in multidrug resistance induced by doxorubicin in colorectal cancer LoVo cells and determine whether telomerase is involved in multidrug resistance.Methods: Adriamycin Method to establish multi-drug resistant model of human colorectal cancer cell line LoVo / Adr; MTT assay was used to identify the drug resistance of drug-resistant LoVo / Adr cells; flow cytometry was used to detect the cycle distribution; RT-PCR was used to detect hTERT mRNA level In the course of induction of drug resistance.Results: The LoVo / Adr cell line was established after continuous culture for 8 months and 67 passages.Meanwhile, LoVo / Adr cell line was established with LoVo / Adr against doxorubicin, vincristine, LoVo / Adr cells compared with LoVo cells, S phase cells decreased, while the G1, G2 phase increased . The low expression of hTERT mRNA in parental LoVo cells was observed in the early stage of doxorubicin induction and then remained high level, which did not increase with the increase of drug resistance.Conclusion: The resistant strain LoVo / Adr is a typical Of multidrug resistance phenotype drug resistance model, telomerase involved in its resistance mechanism.