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P0193 Marsdenia tenacissima extract enhances gefitinib efficacy in non-small-cell lung cancer xenografts
Abstracts / 51 (2015) e1–e36
agents on cancer cells. Cell microencapsulation is a technique by which cells are encapsulated in a semi-permeable microcapsule and live like those in physiological conditions with normal nutritional supply. It is a culturing technique that ranks between monolayer and three-dimensional culture. With microencapsulation, adherent cells grow in a three dimensional manner, and its metabolism and genetic expression also change, similar to solid tumours. Methods: We aimed to establish a cell microencapsulation model using human mucoepidermoid carcinoma cells (MCCs), and assess the cell culture model based on cell growth characteristics, proliferation activity, and protein expression. After MCCs had been isolated and cultured in RPMI-1640 medium, cell growth characteristics, proliferation activity, and protein expression of microencapsulated MCCs and conventional adherent MCCs (as control) were compared. Findings: Results showed MCCs in microcapsules grew in blocks with favourable proliferation activity. On day 3, the cell count in the microencapsulation group was significantly higher than the control group (t = 0.480, p < 0.05). On day 7, cell proliferation had not reached a plateau or declined. Compared with adherent MCCs, microencapsulated MCCs showed significantly higher expression of vascular endothelial growth factor (VEGF) and bFGF (t = 7.617, p < 0.01; t = 6.011, p < 0.01), while TSP-1 was significantly less expressed (t = 12.45, p < 0.001). Interpretation: Microencapsulation culture may help to establish three-dimensional growth of tumour cells in vitro that promote protein expression for angiogenesis. It may be a promising model for functional research of tumour-related genes. In addition, it may be used in studies on biological characteristics of tumour cells and screening of antitumour pharmaceuticals. It will also have prospect in the in situ amplification of stem cells due to convenient inoculation and collection.
http://dx.doi.org/10.1016/j.ejca.2015.06.101
P0191 HISTOGRAM ANALYSIS OF APPARENT DIFFUSION COEFFICIENTS AFTER NEOADJUVANT CHEMOTHERAPY IN BREAST CANCER S.H. Kim *, Y.J. Kim. The Catholic University of Korea, Republic of Korea Background: The purpose of our study was to evaluate change of the apparent diffusion coefficient (ADC) histogram during the neoadjuvant chemotherapy (NAC) in breast cancer, and to compare changes between pathologically verified responders and non-responders. Methods: 62 patients received NAC followed by surgery. We defined responders and non-responders based on 30% reduction in tumour cells using the Miller-Payne Grading System. All patients underwent 3T magnetic resonance (MR) with diffusion-weighted imaging (DWI) before the NAC and after the completion of two cycles of NAC. ADC histogram analysis encompassing the entire tumour was performed and various ADC parameters (mean, minimum, 10th, 25th, 50th, 75th, 90th percentile, maximum ADCs, skewness, and kurtosis) were obtained. Findings: Mean, minimum, 10th, 25th, 50th, and 75th percentile ADCs significantly increased after NAC (p = 0.0129, p = 0.0004, p = 0.0047, p = 0.0002, p = 0.0006, p = 0.0030) and maximum ADC significantly decreased (p = 0.0003). Skewness changed into less positive (p < 0.0001) and kurtosis decreased (p = 0.0168) after NAC. Although there was no statistical significance, mean, minimum, 10th,
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25th, 50th, 75th, and 90th percentile ADCs tended to decrease more in responders compared with non-responders. Interpretation: There is significant change in distribution of ADC after NAC. ADC histogram analysis quantitatively demonstrates the alterations more precisely during the treatment course. http://dx.doi.org/10.1016/j.ejca.2015.06.102
P0192 SENTINEL NODE BIOPSY IN BREAST CANCER USING ONLY METHYLENE BLUE DYE: A PROSPECTIVE STUDY IN A RURAL TERTIARY CARE CENTRE R. Shah *, S. Virani, S. Shah. Kailash Cancer Hospital and Research Centre, Vadodara, India Background: Breast carcinoma is the most common malignancy in women and is the leading cause of death in their middle age. Sentinel lymph node biopsy (SLNB) is a reliable and minimally invasive diagnostic method to determine the regional nodal status in breast cancer and provides accurate staging, such that axillary lymph node dissection can be avoided in negative sentinel node patients. The aim of this study was to assess SLNB, using methylene blue dye, and its accuracy. The complications of using methylene blue dye were also studied. Methods: 138 patients with breast cancer were subjected to SLNB (using methylene blue dye) followed by complete axillary lymph node dissection. The lymph nodes with positive dye were identified. The dye was injected 30 min prior to surgery and the stained lymph nodes were identified during dissection. The haemodynamics of the patients was assessed during and after the procedure. Patients were followed up in the post-operative period, and for the final histopathology report, complications such as wound healing and urine discolouration were assessed. Findings: Of 138 patients with dye injected, 124 (89.85%) patients showed stained lymph nodes. Of 124 patients with positive stain, 67 (54.03%) patients showed evidence of malignancy. Of 67 patients with malignancy-positive lymph nodes, 18 patients had positive sentinel node only. In 49 patients, both sentinel and one or more axillary nodes were positive, whereas in 53 patients both nodal statuses were negative. Four patients had negative sentinel node and positive axillary node. All 14 patients in whom sentinel node could not be identified were negative for cancer in axillary nodes. Seven patients had minor dye-related complications. Interpretation: SLNB with methylene blue dye alone can be considered in breast carcinoma as a reliable, accurate, cost effective, and safe method to detect lymph node status. http://dx.doi.org/10.1016/j.ejca.2015.06.103
P0193 MARSDENIA TENACISSIMA EXTRACT ENHANCES GEFITINIB EFFICACY IN NON-SMALL-CELL LUNG CANCER XENOGRAFTS Shu-Yan Han a,b, Wei Zhao a,c, Hong Sun a,b, Ning Zhou a,b, Fei a Zhou a,b, Guo An a,d, Ping-Ping Li a,b. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University Cancer Hospital & Institute, Beijing, China,
e36
Abstracts / 51 (2015) e1–e36
b Department of Integration of Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing, China, c Department of Cell Biology, Peking University Cancer Hospital & Institute, Beijing, China, d Laboratory Animal Unit, Peking University Cancer Hospital & Institute, Beijing, China
Background: The stem of Marsdenia tenacissima (Roxb.) Wight et Arn. has long been used as a medicine to treat cancer in China. Our previous results showed that M tenacissima extract (MTE) overcomes gefitinib resistance in NSCLC cells. Methods: The present study investigated in vivo anti-tumour activity of MTE combined with gefitinib. H460 (K-RAS mutation) or H1975 cells (T790M mutation) were subcutaneously inoculated into nude mice. Tumour volume and body weight were measured during the experiment. The resected tumours were weighed after animals were sacrificed. Cellular proliferation and apoptosis in xenografts tumour tissue were assessed. EGFR downstream pathways and c-Met expression was evaluated by western blotting. In accordance with the previous in vitro study, MTE at low dose (5 g/kg) was chosen to assess whether it can restore gefitinib sensitivity in vivo.
Findings: MTE enhanced gefitinib efficacy in resistant H460 and H1975 xenografts. The combination significantly inhibited tumour proliferation and induced cell apoptosis in both resistant NSCLC xenografts. Constitutive activation of PI3K/Akt and MEK/ERK pathway is related to EGFR-TKI resistance. Accordingly, phosphorylation of PI3K/Akt/mTOR and ERK1/2 was suppressed after the combined treatment. Simultaneously, the cross-talk between c-Met and EGFR was also lowered in the presence of MTE combined with gefitinib. Interpretation: This study provides in vivo evidence of MTE enhancing gefitinib efficacy in resistant NSCLC xenografts, and suggests that the combination of MTE and gefitinib could be a promising approach against NSCLC.
http://dx.doi.org/10.1016/j.ejca.2015.06.104
agents on cancer cells. Cell microencapsulation is a technique by which cells are encapsulated in a semi-permeable microcapsule and live like those in physiological conditions with normal nutritional supply. It is a culturing technique that ranks between monolayer and three-dimensional culture. With microencapsulation, adherent cells grow in a three dimensional manner, and its metabolism and genetic expression also change, similar to solid tumours. Methods: We aimed to establish a cell microencapsulation model using human mucoepidermoid carcinoma cells (MCCs), and assess the cell culture model based on cell growth characteristics, proliferation activity, and protein expression. After MCCs had been isolated and cultured in RPMI-1640 medium, cell growth characteristics, proliferation activity, and protein expression of microencapsulated MCCs and conventional adherent MCCs (as control) were compared. Findings: Results showed MCCs in microcapsules grew in blocks with favourable proliferation activity. On day 3, the cell count in the microencapsulation group was significantly higher than the control group (t = 0.480, p < 0.05). On day 7, cell proliferation had not reached a plateau or declined. Compared with adherent MCCs, microencapsulated MCCs showed significantly higher expression of vascular endothelial growth factor (VEGF) and bFGF (t = 7.617, p < 0.01; t = 6.011, p < 0.01), while TSP-1 was significantly less expressed (t = 12.45, p < 0.001). Interpretation: Microencapsulation culture may help to establish three-dimensional growth of tumour cells in vitro that promote protein expression for angiogenesis. It may be a promising model for functional research of tumour-related genes. In addition, it may be used in studies on biological characteristics of tumour cells and screening of antitumour pharmaceuticals. It will also have prospect in the in situ amplification of stem cells due to convenient inoculation and collection.
http://dx.doi.org/10.1016/j.ejca.2015.06.101
P0191 HISTOGRAM ANALYSIS OF APPARENT DIFFUSION COEFFICIENTS AFTER NEOADJUVANT CHEMOTHERAPY IN BREAST CANCER S.H. Kim *, Y.J. Kim. The Catholic University of Korea, Republic of Korea Background: The purpose of our study was to evaluate change of the apparent diffusion coefficient (ADC) histogram during the neoadjuvant chemotherapy (NAC) in breast cancer, and to compare changes between pathologically verified responders and non-responders. Methods: 62 patients received NAC followed by surgery. We defined responders and non-responders based on 30% reduction in tumour cells using the Miller-Payne Grading System. All patients underwent 3T magnetic resonance (MR) with diffusion-weighted imaging (DWI) before the NAC and after the completion of two cycles of NAC. ADC histogram analysis encompassing the entire tumour was performed and various ADC parameters (mean, minimum, 10th, 25th, 50th, 75th, 90th percentile, maximum ADCs, skewness, and kurtosis) were obtained. Findings: Mean, minimum, 10th, 25th, 50th, and 75th percentile ADCs significantly increased after NAC (p = 0.0129, p = 0.0004, p = 0.0047, p = 0.0002, p = 0.0006, p = 0.0030) and maximum ADC significantly decreased (p = 0.0003). Skewness changed into less positive (p < 0.0001) and kurtosis decreased (p = 0.0168) after NAC. Although there was no statistical significance, mean, minimum, 10th,
e35
25th, 50th, 75th, and 90th percentile ADCs tended to decrease more in responders compared with non-responders. Interpretation: There is significant change in distribution of ADC after NAC. ADC histogram analysis quantitatively demonstrates the alterations more precisely during the treatment course. http://dx.doi.org/10.1016/j.ejca.2015.06.102
P0192 SENTINEL NODE BIOPSY IN BREAST CANCER USING ONLY METHYLENE BLUE DYE: A PROSPECTIVE STUDY IN A RURAL TERTIARY CARE CENTRE R. Shah *, S. Virani, S. Shah. Kailash Cancer Hospital and Research Centre, Vadodara, India Background: Breast carcinoma is the most common malignancy in women and is the leading cause of death in their middle age. Sentinel lymph node biopsy (SLNB) is a reliable and minimally invasive diagnostic method to determine the regional nodal status in breast cancer and provides accurate staging, such that axillary lymph node dissection can be avoided in negative sentinel node patients. The aim of this study was to assess SLNB, using methylene blue dye, and its accuracy. The complications of using methylene blue dye were also studied. Methods: 138 patients with breast cancer were subjected to SLNB (using methylene blue dye) followed by complete axillary lymph node dissection. The lymph nodes with positive dye were identified. The dye was injected 30 min prior to surgery and the stained lymph nodes were identified during dissection. The haemodynamics of the patients was assessed during and after the procedure. Patients were followed up in the post-operative period, and for the final histopathology report, complications such as wound healing and urine discolouration were assessed. Findings: Of 138 patients with dye injected, 124 (89.85%) patients showed stained lymph nodes. Of 124 patients with positive stain, 67 (54.03%) patients showed evidence of malignancy. Of 67 patients with malignancy-positive lymph nodes, 18 patients had positive sentinel node only. In 49 patients, both sentinel and one or more axillary nodes were positive, whereas in 53 patients both nodal statuses were negative. Four patients had negative sentinel node and positive axillary node. All 14 patients in whom sentinel node could not be identified were negative for cancer in axillary nodes. Seven patients had minor dye-related complications. Interpretation: SLNB with methylene blue dye alone can be considered in breast carcinoma as a reliable, accurate, cost effective, and safe method to detect lymph node status. http://dx.doi.org/10.1016/j.ejca.2015.06.103
P0193 MARSDENIA TENACISSIMA EXTRACT ENHANCES GEFITINIB EFFICACY IN NON-SMALL-CELL LUNG CANCER XENOGRAFTS Shu-Yan Han a,b, Wei Zhao a,c, Hong Sun a,b, Ning Zhou a,b, Fei a Zhou a,b, Guo An a,d, Ping-Ping Li a,b. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University Cancer Hospital & Institute, Beijing, China,
e36
Abstracts / 51 (2015) e1–e36
b Department of Integration of Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing, China, c Department of Cell Biology, Peking University Cancer Hospital & Institute, Beijing, China, d Laboratory Animal Unit, Peking University Cancer Hospital & Institute, Beijing, China
Background: The stem of Marsdenia tenacissima (Roxb.) Wight et Arn. has long been used as a medicine to treat cancer in China. Our previous results showed that M tenacissima extract (MTE) overcomes gefitinib resistance in NSCLC cells. Methods: The present study investigated in vivo anti-tumour activity of MTE combined with gefitinib. H460 (K-RAS mutation) or H1975 cells (T790M mutation) were subcutaneously inoculated into nude mice. Tumour volume and body weight were measured during the experiment. The resected tumours were weighed after animals were sacrificed. Cellular proliferation and apoptosis in xenografts tumour tissue were assessed. EGFR downstream pathways and c-Met expression was evaluated by western blotting. In accordance with the previous in vitro study, MTE at low dose (5 g/kg) was chosen to assess whether it can restore gefitinib sensitivity in vivo.
Findings: MTE enhanced gefitinib efficacy in resistant H460 and H1975 xenografts. The combination significantly inhibited tumour proliferation and induced cell apoptosis in both resistant NSCLC xenografts. Constitutive activation of PI3K/Akt and MEK/ERK pathway is related to EGFR-TKI resistance. Accordingly, phosphorylation of PI3K/Akt/mTOR and ERK1/2 was suppressed after the combined treatment. Simultaneously, the cross-talk between c-Met and EGFR was also lowered in the presence of MTE combined with gefitinib. Interpretation: This study provides in vivo evidence of MTE enhancing gefitinib efficacy in resistant NSCLC xenografts, and suggests that the combination of MTE and gefitinib could be a promising approach against NSCLC.
http://dx.doi.org/10.1016/j.ejca.2015.06.104