个体价值认同的社会主义核心价值观培育探析

目的 建立人结直肠癌耐奥沙利铂(L-OHP)细胞株,检测其多药耐药性并初步探讨可能的耐药机制。 方法 采用药物浓度梯度递增诱导法建立人结直肠癌耐L-OHP细胞株HCT-8/L-OHP。MTT法鉴定药敏性,并检测耐药细胞株的多药耐药性、生长曲线和细胞群体倍增时间。用光镜、电镜、流式细胞术等方法观察亲本细胞和耐药细胞的形态、周期及细胞凋亡。通过实时荧光定量PCR和Western-blot检测HOXB8基因和ABCC1(MRP or MRP1),ERCC1,P-gp,GST-π的mRNA及蛋白表达水平。 结果 建成了肠癌耐药细胞模型HCT-8/L-OHP,半抑制浓度(IC50)为62.33 μmol/L,耐药指数(RI)为10.78。耐药株细胞形态发生变化,代谢率降低,生物活力减弱。HCT-8/L-OHP细胞株具有耐药性,细胞增殖减慢。HCT-8/L-OHP细胞在G₂/M期的比例增多,在S期及G₀/G₁期的比例减少,相较于亲本细胞株,差别具有统计学意义(P<0.01); HCT-8/L-OHP细胞凋亡率低于HCT-8细胞。ABCC1(MRP or MRP1),ABCB1(P-gp)等耐药相关基因在耐药细胞株中高表达; HOXB8基因在HCT-8/L-OHP细胞中表达,较亲本细胞株表达明显升高,差别具有统计学意义(P<0.01)。 结论 成功构建了肠癌耐药细胞株HCT-8/L-OHP,耐药的分子机制可能是ABCC1(MRP or MRP1)及ABCB1(P-gp)等基因的表达升高。HOXB8基因过表达可能与人结直肠癌对L-OHP耐药的机制相关。

Establishment of Oxaliplatin-resistance Colorectal Cancer Cell Lines HCT-8/L-OHP and an Explorative Analysis of HOXB8 Overexpression in HCT-8/L-OHP

ABSTRACT: Objective To establish and authenticate an oxaliplatin(L-OHP)resistant human colon cancer cell line,and to explore the mechanism of L-OHP resistance. Methods Resistance to oxaliplatin was induced in HCT-8(HCT-8/L-OHP)colorectal adenocarcinoma cell lines by exposing cells to increasing concentrations of L-OHP. MTT, Flow cytometry(FCM), RT-qPCR, and Western-blot were used to evaluate the resistance to L-OHP. Results Built the stable HCT-8/L-OHP cell model. The drug resistance index(RI)of HCT-8/L-OHP cell line to the L-OHP, 5-FU, CPT-11, VCR, and CDDP were 10.87, 5.35, 3.23, 1.28, and 6.64, respectively. Increased expression of multi-drug resistant genes ABCC1, ABCB1, GSTP1, and ERCC1 were detected in HCT-8/L-OHP cell line. Conclusion We successfully established the L-OHP resistant HCT-8/L-OHP cell line. In addition, the elevated expression of ABCC1, ABCB1 and ERCC1 genes may be associated with molecular mechanism in oxaliplatin resistance. And HOXB8 overexpression is involved in L-OHP resistance mechanism of human colon cancer.