Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies / 细胞与分子免疫学杂志

Objective To prepare and identify rabbit anti-cyclin dependent kinase 6 (CDK6) antibody. Methods The recombinant pET21a (+)/CDK6 was successfully constructed, then the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells and was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for protein expression, which was detected by SDS-PAGE and Western blot analysis. The expressed protein was purified by nickel-chelating nitrilotriacetic acid (Ni-NTA) agarose and then analyzed by SDS-PAGE. Japanese white rabbits were immunized with purified CDK6 protein for many times every two weeks. The blood was collected at 0, 2, 4 and 6 weeks after immunization, and serum was separated from blood. The titer was detected by indirect ELISA. Western blot analysis, immunofluorescence assay and immunohistochemistry were employed to determine the specificity. Results High purity CDK6 protein and high specificity of rabbit anti-CDK6 antibody were successfully prepared. The titer of CDK6 rabbit serum antibody reached 1:30 000 after immunization, which could specifically recognize the CDK6 protein expressed in cervical cancer cell line and cervical cancer tissues. Conclusion The high titer and specificity of rabbit anti-CDK6 antibody is successfully prepared.

Pemetrexed and platinum as first-line chemotherapy for advanced lung adenocarcinoma:an observation of recent therapeutic effects and a safety profile / 中国肿瘤临床

Objective:The recent therapeutic effects and safety profile of pemetrexed plus cisplatin as first-line chemotherapy in advanced lung adenocarcinoma were evaluated. Methods:A total of 41 chemotherapy-naive locally advanced or metastatic non-small cell lung cancer patients, who were diagnosed with adenocarcinoma by pathological examination, were included. All patients received 500 mg/m2 pemetrexed on day 1. A total dose of 75 mg/m2 cisplatin was divided into three daily doses administered over 3 d. Treat-ments were repeated every three weeks. The therapeutic efficiency and safety profile were evaluated every two treatment cycles. Re-sults:All 41 patients were eligible for the evaluation of therapeutic efficiency. Seventeen patients showed partial response, 20 patients showed stable disease, and four patients showed disease progression. The overall response rate (ORR) was 41.5%and the disease con-trol rate (DCR) was 90.3%. Subgroup analysis showed that the ORR in males was significantly higher than that in females (63.1%vs. 22.7%, P=0.009). The median progression-free survival of all patients was 11.0 months, whereas the median overall survival was 12.6 months. The intracranial lesions of six patients with brain metastases were well controlled after chemotherapy and palliative whole brain radiotherapy. The ORR and DCR for these patients were 83.3%and 100.0%, respectively. In terms of safety, only 4.8%of the pa-tients showed gradesⅢandⅣneutropenia, nausea, or vomiting. Conclusion:Pemetrexed plus cisplatin is an effective and well-tolerat-ed regimen as first-line therapy for patients with lung adenocarcinoma. According to subgroup analysis, efficacy in males was signifi-cantly better than that in females. Good efficacy was observed in patients with brain metastases treated with chemotherapy and pallia-tive whole brain radiotherapy. No additional adverse effects were observed.