The biological clock gene BMAL1 inhibits the proliferation, migration and invasion of radiation-resistant nasopharyngeal carcinoma cell line 5-8FR by regulating PI3K/Akt/MMP-2/9 signaling pathway / 中华放射肿瘤学杂志

Objective:To investigate the effect of BMAL1 gene on the proliferation, migration and invasion ability of radiation-resistant nasopharyngeal carcinoma cell line (5-8FR) and the molecular mechanism. Methods:A multi-target click model was constructed for radiation-resistant nasopharyngeal carcinoma cell line 5-8FR by low-dose fractionated irradiation, and the results of clone formation assay were used to fit the multi-target click model and calculate the sensitization ratio of radiotherapy. The expression levels of PI3K/Akt/MMP-2/9 signaling pathway-related proteins in 5-8FR and control 5-8F cell lines were detected by Western blot. The overexpression and knockdown vectors of BMAL1 gene were constructed and transfected with 5-8F and 5-8F cell lines, respectively. The BMAL1 gene overexpression (pcDNA-BMAL1) and its control (pcDNA) and interference (BMAL1-shRNA) and control (con-shRNA) cell lines were stably transfected with nasopharyngeal carcinoma cell line 5-8F and radiation-resistant cell line 5-8FR, respectively. Western blot was performed to verify the infection efficiency and detect the changes of PI3K/Akt/MMP-2/9 signaling pathway-related proteins after overexpression or interference of BMAL1 gene in both groups of cells. CCK-8 assay, cell scratch test and Transwell chamber test were conducted to investigate the proliferation, migration and invasion capabilities of 5-8FR cell line after overexpression or interference of BMAL1 gene. Results:BMAL1 gene expression was down-regulated, and those of PI3K/Akt pathway proteins and downstream related molecules of MMP-2 and MMP-9 were up-regulated, and TIMP-2 and TIMP-1 expression was down-regulated in nasopharyngeal carcinoma radiation-resistant cell lines. Overexpression of BMAL1 gene inhibited the expression of PI3K/Akt pathway proteins and downstream related molecules of MMP-2 and MMP-9, promoted the expression of TIMP-2 and TIMP-1, and inhibited the proliferation, migration and invasion capabilities of radiation-resistant nasopharyngeal carcinoma cells, while interference with BMAL1 gene yielded the opposite results. Conclusions:BMAL1 gene can reverse the expression of PI3K/Akt/MMP-2/9 signaling pathway-related proteins in radiation-resistant nasopharyngeal carcinoma cell lines and inhibit the proliferation, migration and invasion capabilities of radiation-resistant nasopharyngeal carcinoma cell lines.

A clinical study of perfusion weighted magnetic resonance imaging in recombinant human endostatin combined with chemotherapy and radiotherapy for locally advanced nasopharyngeal carcinoma / 中华放射医学与防护杂志

Objective To assess the antiangiogenic role of recombinant human endostatin combined with chemoradiotherapy and the capacity,and to explore the early tumor response as measured by comparing the change of MRI perfusion parameter.Methods From May 2012 to March 2013,22 locally advanced nasopharyngeal carcinoma patients who received recombinant human endostatin combined with chemoradiotherapy following induction chemotherapy,were included in the prospective study group.The other 25 patients,who received chemoradiotherapy following induction chemotherapy alone in the same period,were included in the control group.The perfusion parameters including blood volume(BV),blood flux(BF),mean transit time (MTT) were obtained by carrying out MR perfusion scanning at 3 time points:before induction chemotherapy,after induction chemotherapy,the end of concurrent chemoradiotherapy.Results Compared with before induction chemotherapy,the perfusion parameters including BV and BF obviously decreased in the study group (F =3.05,3.85,P ＜ 0.05).The parameter of MTT had no obviously change in the study group(P ＞0.05).In the control group,the change of BV,BF and MTT of nasopharyngeal lesions area during the treatment showed no significant difference (P ＞ 0.05).To make comparison between the two groups,at the end of concurrent chemoradiotherapy,BF of nasopharyngeal lesions area in the study group was 0.72 ± 0.56 and 1.92 ± 1.26 in the control group,the former showing significantly declined results (t =-3.056,P =0.012).Conclusions Recombinant human endostatin might be a good indicator of local tumor microvascular changes and the treatment-related toxicity could be tolerated.Magnetic resonance perfusion imaging maybe assessed the capacity of anti-angiogenesis therapy to induce early tumor response.Clinical trial registration Chinese clinical trial registry,ChiCRTONRC-12002394.

Phase II clinical trial of two different modes of administration of the induction chemotherapy for locally advanced nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare the therapeutic effects, toxic side effects and influence on the immune function in patients treated with TPF [docetaxel (DOC) + cisplatin (DDP) + 5-fluorouracil (5-Fu)] induction chronochemotherapy and conventional chemotherapy for locally advanced nasopharyngeal (NPC).</p><p><b>METHODS</b>Seventy patients with locally advanced nasopharyngeal carcinoma were treated in our department at their first visit from April 2013 to December 2013. They were divided randomly into two groups: the chronochemotherapy group (38 patients) and conventional chemotherapy group (32 patients). All of the patients were treated with TPF regimen with 2 cycles of induction chemotherapy in a 21-28-days/cycle. The chronochemotherapy group: DOC: 75 mg/m2, i. v. gtt, d1 (03: 30-04: 30); DDP: 75 mg/m2, 10 am-10 pm, c.i.v, d1-d5; 5-Fu: 750 mg·m(-2)·d(-1), 10 pm-10 am, c. i.v., d1-d5, both chemotherapies were administered by intravenous infusion using an automatic electric pump. The conventional chemotherapy group: Both DOC and DDP were administered intravenously at a dose of 75 mg/m2 on d1. 5-Fu was given at a dose of 750 mg/m2 for 24 hours from d1-d5 with continuous infusion in a total of 120 hours. In this procedure, prescribing the conventional intravenous infusion, intensity modulated radiation therapy was used after the induction chemotherapy. The prescribed nasopharyngeal lesion dose (GTVnx) was 69.96 Gy/33 fractions for the T1-T2 nasopharygeal cancer, while 73.92 Gy/33 fractions nasopharynx lesion dose (GTVnx) for the T3-T4 nasopharyngeal cancer. The planning target volume (PTV) of positive lymph node (PTVnd) dose was 69.96 Gy/33 fractions. Concurrent chemoradiotherapy: cisplatin 100 mg/m2, i. v. gtt. d1-d2, and there were two cycles in total and 21 days each cycle.</p><p><b>RESULTS</b>Sixty-six patients were evaluable for the response assessment. There were 36 patients in the chronochemotherapy group and 30 patients in the conventional chemotherapy group. After the induction chemotherapy, no CR case was found in both of the two groups. The PR was 80.6% in the chronochemotherapy group and 50.0% in the conventional chemotherapy group (P=0.009). After concurrent chemoradiotherapy, the CR rate in the chronocheotherapy group was 45.5%, significantly higher than 20.7% in the conventional chemotherapy group (P=0.040). Secondly, the incidence rates of adverse reactions including bone marrow suppression, nausea, vomiting, diarrhea, constipation, oral mucositis, fatigue, anorexia in the chrono-chemotherapy group were significantly lower than that in the conventional group (P<0.05 for all). Finally, compared the two groups, the CD4+ /CD8+ ratio was significantly lower in the chronochemotherapy group than that in the conventional chemotherapy group (P<0.05). The lymphocytes CD19+ and CD4+/CD8+ were decreased and CD3+, CD4+, CD8+, CD16++CD56+ were increased in the chronochemotherapy group, while only CD3+ and CD8+ were increased in the conventional chemotherapy group.</p><p><b>CONCLUSIONS</b>Compared with the conventional chemotherapy, the chronochemotherapy may be more favorable in the treatment of NPC, with a better therapeutic effects and effectiveness than that of conventional chemotherapy after induction chemotherapy, with less side effects, and can improve the immune function in the patients.</p>

Clinical results of recombinant human endostatin combined with chemoradiotherapy for locally advanced nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare the short-term efficacy and observe the tolerability and safety of recombinant human endostatin combined with induction chemotherapy followed by chemoradiotherapy for locally advanced nasopharyngeal carcinoma.</p><p><b>METHODS</b>Fifty-three patients with locally advanced nasopharyngeal carcinoma, who received recombinant human endostatin combined with induction chemotherapy followed by chemoradiotherapy, treated in our department from December 2011 to March 2013 were included in the study group of this study. Another 48 patients, who received induction chemotherapy followed by chemoradiotherapy alone in the same period, were chosen as a control group. The short-term outcome, overall survival (OS), progression-free survival (PFS), and acute side effects of the two groups were compared.</p><p><b>RESULTS</b>The complete remission rates of nasopharyngeal tumor in the study and control groups were 77.4% and 72.9%, respectively (P=0.154). The complete remission rates of patients with and without cervical lymph node metastasis were 75.5% and 62.6%, respectively, showing a significant difference (P=0.037). The 2-year OS, PFS, and DMFS rates for the study group were 82.3%, 77.2%, and 82.2%, respectively, versus 87.2%, 84.3% and 84.2% for the control group, showing a non-significant differences between the two groups (P=0.938, P=0.551, and P=0.725).</p><p><b>CONCLUSIONS</b>The short-term results of recombinant human endostatin (Endostar) combined with induction chemotherapy followed by concurrent chemoradiotherapy in the treatment of locally advanced nasopharyngeal carcinoma are slightly better than that of induction chemotherapy followed by concurrent chemoradiotherapy alone, with tolerable treatment-related toxicity and no more side effects.</p>

Clinical study of chrono-chemotherapy in treating nasopharyngeal carcinoma patients with distant metastasis at preliminary diagnosis / 中国肿瘤临床

Objective:To investigate the outcomes of the regimen with docetaxel, cisplatin, and 5-fluorouracil (TPF regimen) in chrono-chemotherapy, and evaluate the feasibility of reducing the toxicity and immunological damage in nasopharyngeal carcinoma (NPC) patients with distant metastasis at preliminary diagnosis, then to compare the advantages and disadvantages between chrono-che-motherapy and traditional chemotherapy. Methods:A total of 46 NPC patients with distant metastasis at preliminary diagnosis (UICC 2010 stage IVc) were enrolled in this study. These NPC patients were randomly divided into chrono-chemotherapy and conventional chemotherapy groups, with 23 cases for each group. TPF neo-adjuvant chemotherapy was conducted in both groups for two cycles, with 21 days to 28 days for each cycle. The following regimen was used for the chrono-chemotherapy group:docetaxel 75 mg/m2, infu-sion, d1;cisplatin 75 mg/m2, 10:00 a.m.-10:00 p.m., continuous infusion, d1-d5;and fluorouracil 750 mg/(m2 · d), 10:00 p.m.-10:00 a. m., continuous intravenous infusion, d1-d5. The following regimen was used for the conventional chemotherapy group:docetaxel 75 mg/m2, infusion, d1;cisplatin 75 mg/m2, infusion, d1;and fluorouracil 750 mg/(m2· d), continuous infusion, d1-d5, 120 h. Patients who obtained therapeutic efficacy via induction chemotherapy were provided with intensity-modulated radiotherapy as a concurrent radio-therapy and chemotherapy (DDP 100 mg/m2, infusion, d1-d2, with 21 days each cycle and a total of two courses). One month after con-current chemoradiation, an adjuvant chemotherapy with the same regimen as the induction chemotherapy was employed for a total of two courses. Acute and late toxicities were graded in accordance with the Common Terminology Criteria for Adverse Events v3.0 scor-ing. Tumor response was evaluated using the 2000 Response Evaluation Criteria in Solid Tumors. The effective rates included complete and partial responses. Relevant data were analyzed by SPSS16.0 statistical software. Results:More emesis was observed at Grade 2 or above in the conventional chemotherapy group than in the chrono-chemotherapy group, with statistical significance between the two groups (P=0.035). After chemotherapy, the value of CD4/CD8 increased in the chrono-chemotherapy group and decreased in the con-ventional chemotherapy group, with statistical significance between the two groups (P=0.033). Conclusion:The proposed chrono-che-motherapy outperforms conventional chemotherapy in reducing the occurrence of severe vomiting. This chrono-chemotherapy may be advantageous in reducing severe bone marrow depression and may play a positive role in the immune function of NPC patients.

PhaseⅡclinical trial of induction chrono-chemotherapy followed by concurrent chemoradiation in loco-regionally advanced nasopharyngeal carcinoma / 中国肿瘤临床

Objective:The present study aimed to investigate the short-term efficacy and adverse effects of induction chrono-che-motherapy including docetaxe1 (TXT), cisplatin (DDP), and 5 fluorouraci1 (5-FU) followed by concomitant chemoradiotherapy in lo-co-regionally advanced nasopharyngeal carcinoma (NPC). Methods:Newly diagnosed locally advanced (Ⅲ~Ⅳb) NPC patients were enrolled in this study. All patients received three cycles of TPF regimen. The TPF chemotherapy regimen was administered as follows:TXT, 75 mg/m2, i.v. infusion, d1; DDP, 75 mg/m2, bolus infusion from 10:00 to 22:00, d1-5; and 5-FU 750 mg/m2/d bolus infusion from 22:00 to 10:00, d1-5, with 21 days each cycle, followed by concomitant IMRT and chemotherapy (paclitaxel 135 mg/m2 i.v. infu-sion, with 21 days each cycle and a total of 2 courses). Acute and late toxicities were graded according to the Common Terminology Cri-teria for Adverse Events v3.0 scoring criteria. Tumor response was evaluated using 2000 Response Evaluation Criteria in Solid Tumors criteria. Results:The CR and PR rates of induction chemotherapy were 23.8%and 68.6%, respectively;whereas the CR and PR rates of the combined modality treatment were 64.8%and 31.4%, respectively. Two-year overall survival rate was 91.4%, two-year progres-sion free survival rate was 87.0%, and two-year distant metastasis-free survival rate was 88.4%. The main side effects from induction chemotherapy include an over grade 3 granulocytopenia of 28.6%. Major toxicity from concurrent chemo-radiotherapy was oral mucosi-tis (81.0%);grade 3 to 4 oral mucositis was 16%. No treatment-related deaths occurred in this study. Conclusion:Induction chrono-che-motherapy using TPF followed by concurrent chemoradiotherapy of paclitaxel is a well-tolerated treatment with short-term efficacy and severity for locally advanced NPC. Further follow-up is required to assess the late effects and long-term efficacy.