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目的观察化疗药物对原代胃癌细胞的体外杀伤效应,并探讨其与胃癌组织中端粒酶逆转录酶(hTERT)mRNA表达的关系。方法将68例新鲜胃癌组织制备单细胞悬液,台盼蓝染色计数活细胞,分别加入紫杉醇(TZX)、阿霉素(ADM)、顺铂(CDDP)、5-氟尿嘧啶(5-Fu)和丝裂霉素(MMC)培养48 h。四甲基偶氮唑盐(MTT)法观察药物作用后肿瘤细胞活力及代谢活性变化。用原位杂交技术检测hTERT mRNA的表达。结果不同个体的肿瘤细胞对化疗药物敏感性不同,与肿瘤分化有关,TAX、CDDP、5-Fu、ADM和MMC的胃癌细胞抑制率分别为(40.6±6.9)%、(38.4±7.8)%、(38.9±9.2)%、(31.6±8.5)%和(28.9±9.8)%。全组hTERT mRNA表达阳性率为90.0%,hTERT mRNA阳性者对5-Fu和ADM有较强的耐药性。结论hTERT mRNA的高表达可能是胃癌原发性耐药的标志之一,MTT比色法体外药敏实验有助于筛选胃癌的个体化有效化疗药物。
Objective To observe the killing effect of chemotherapeutic drugs on primary gastric cancer cells in vitro and to explore its relationship with telomerase reverse transcriptase (hTERT) mRNA expression in gastric cancer. Methods Sixty-eight fresh gastric cancer tissues were prepared for single cell suspension and viable cells were counted by trypan blue staining. TZX, ADM, CDDP, 5-Fu and Mitomycin (MMC) for 48 h. The MTT assay was used to observe the change of tumor cell viability and metabolic activity after drug treatment. Detection of hTERT mRNA expression by in situ hybridization. Results The sensitivity of different tumor cells to chemotherapeutic agents was different, which was related to tumor differentiation. The inhibitory rates of gastric cancer cells TAX, CDDP, 5-Fu, ADM and MMC were (40.6 ± 6.9)% and (38.4 ± 7.8)%, (38.9 ± 9.2)%, (31.6 ± 8.5)% and (28.9 ± 9.8)%, respectively. The positive rate of hTERT mRNA expression in the whole group was 90.0%. The positive hTERT mRNA was highly resistant to 5-Fu and ADM. Conclusion High expression of hTERT mRNA may be one of the markers of primary drug resistance in gastric cancer. MTT assay in vitro may be useful in screening individual and effective chemotherapeutic agents for gastric cancer.