目的 构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型。方法 采用低浓度加量持续诱导法,诱导吉西他滨耐药乳腺癌细胞4T1耐药株,命名为4T1/Gem;CCK-8法测定4T1与4T1/Gem细胞的增殖抑制率,计算耐药指数; Western blot法检测细胞P-gp蛋白表达;B超引导下注射4T1/Gem细胞悬液诱导裸鼠肝脏成瘤;HE染色观察肿瘤组织病理情况,免疫组化法检测瘤组织ER、PR、HER2、Ki-67和P-gp蛋白的表达。结果 经过14个月的诱导成功建立4T1/Gem细胞株,可在含40 μg/mL的Gem培养液中稳定生长。4T1/Gem细胞耐药指数为4T1细胞的788.547倍。与亲代相比,4T1/Gem处于G1期和G2期的细胞增加,S期细胞减少;上调P-gp蛋白的表达。4T1/Gem细胞成功建立裸鼠乳腺癌肝转移模型,瘤组织中ER、PR、HER2蛋白阴性表达,Ki-67阳性10％和P-gp蛋白阳性表达。结论 成功构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型,为开发治疗乳腺癌肝转移化疗耐药的药物提供实验基础。

Objective To construct a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem) and establish a nude mouse model of breast cancer with hepatic metastatic. Methods A gemcitabine-resistant variant of the breast cancer 4T1 cell line was induced by gradually increasing the concentration of gemcitabine; this variant is referred to in this study as 4T1/Gem. The proliferation suppression rates of 4T1 and 4T1/Gem cells were determined by using the CCK-8 essay to evaluate the drug resistance indices of the cell lines. Western blot analysis was used to detect P-gp protein expression. Under ultrasonography, a 4T1/Gem cell suspension was injected into nude mice to induce liver tumors. H&E staining was used to observe tumor pathology, and immunohistochemistry was used to detect the expression of ER, PR, HER-2, Ki-67, and P-gp. Results After 14 months of induction, a 4T1/Gem cell line is established successfully. The cell line can grow stably in culture liquid containing 40 μg/ml gemcitabine. The drug resistance index of 4T1/Gem is 788.547. Compared with the 4T1 cell line, the 4T1/Gem cell line can upregulate P-gp protein expression and successfully establish a nude mouse model of breast cancer with hepatic metastatic. ER, PR, and HER-2 proteins exhibit negative expression in the tumor tissue. The positive expression of P-gp and 10% of Ki-67 proteins is also observed. Conclusion This study successfully constructs a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem)and establishes a nude mouse model of breast cancer with hepatic metastatic, thereby providing an experimental basis for developing and treating a drug-resistant variant of breast cancer.

目的 观察吉西他滨联合萘达铂治疗铂类敏感复发性卵巢癌的近期疗效与不良反应。方法 回顾性分析潍坊市人民医院2013年1月—2014年6月治疗的60例复发性卵巢癌病例,分为吉西他滨联合萘达铂(GN)方案化疗组30例和吉西他滨联合卡铂(GC)方案化疗组30例。GN方案组,吉西他滨1.0 g/m²,d1、8;萘达铂80 mg/m²,d1,21d为1周期;GC方案组,吉西他滨1.0 g/m²,d1、8;卡铂按AUC=5计算,d1,21 d为1周期。结果 GN方案组近期有效率56.0%,GC方案组近期有效率60.0%,两组比较差异无统计学意义(χ² =0.069,P=0.793)。两组最常见的毒性反应均是骨髓抑制,GC组骨髓抑制发生率较GN组骨髓抑制发生率稍高,但两组比较差异无统计学意义(P>0.05)。结论 两种方案治疗铂类敏感型复发性卵巢癌疗效无统计学差异,GN方案组不良反应较轻。

Objective To evaluate the clinical effect of GN chemotherapy protocol and GC chemotherapy protocol treatment of recurrent ovarian cancers. Methods We retrospectively analyzed the data of 60 patients with recurrent ovarian cancers in our hospital from January 2013 to June 2014. Divided into gemcitabine and nedaplatin (GN)chemotherapy group and Gemcitabine and carboplatin(GC)chemotherapy group, 30 patients in each group.Patients in GP protocol group were given Gemcitabine 1.0 g/m²,d1,8;and naphthalene(80 mg/m²,d1),21d was a period of treatment; Patients in GC protocol group were given gemcitabine 1.0 g/m²,d1,8;and carboplatin AUC=5 by calculation,21d was a period of treatment. Results For patients in GN protocol group,the short term response rate was 56.0%.For patients in GC protocol group,the short term response rate was 60.0%,the difference was not statistically significant(χ²=0.069,P=0.793). Two of the most common toxicities were myelosuppression, incidence of myelosuppression GC was slightly higher than GN, and the difference was not statistically significant. Conclusion The therapeutic effects of two chemotherapy protocols have no statistically significant difference in treatment of platinum sensitive recurrent ovarian cancer. Toxicity of GN group is light.