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SLC40A1 Sensitized Human Ovarian Cancer Cells to Cisplatin by Blocking Autophagy
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Abstracts / Journal of Minimally Invasive Gynecology 24 (2017) S1–S201 Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:45 AM – STATION G
Influence of Race/Ethnicity on Route of Hysterectomy and Inpatient Surgical Complications Bougie O,1 Singh SS,1 McCarthy EP2. 1Ottawa Hospital Research Institute, University of Ottawa, Ottawa, Ontario, Canada; 2Harvard T.H. Chan School of Public Health, Boston, Massachusetts Study Objective: To examine the association between race and (1) route of hysterectomy and (2) risk of inpatient surgical complications. Design: A cross-sectional analysis of women who underwent hysterectomy during 2009–2013 using the Nationwide Inpatient Sample. Setting: The Nationwide Inpatient Sample (NIS) is 20% stratified sample of all patient discharges from acute care hospitals in the United States designed to be nationally representative. Patients: We identifed 114,719 women age 18 and older who underwent elective hysterectomy for benign indications using International Classification of Diseases (ICD-9-CM) procedure and diagnostic codes. Intervention: Our main factor of interest was race/ethnicity, categorized as: White, Black, Hispanic, Other or missing. Measurements and Main Results: Compared to white women, black women had 0.55 (95%CI: 0.52–0.59) odds of receiving minimally invasive hysterectomy, adjusted for age, Median household income national quartile for patient ZIP Code, primary payer, hospital location/teaching status, Elixhauser comorbidity index, and indication for surgery. This finding remained consistent across household income quartile for patient ZIP Code, primary payer and diagnosis of fibroids. In our sample of women, 6091 experienced a complication, representing 30,455 women nationwide. Compared to white women, the odds of black women experiencing a surgical complication was 1.03(95% CI: 0.93– 1.13) after adjusting for age, median household income national quartile for patient ZIP Code, primary payer, hospital location/teaching status, Elixhauser comorbidity index, and route of surgery. This finding remained consistent across household income quartile for patient ZIP Code, primary payer and route of hysterectomy. Conclusion: Among women who underwent hysterectomy for benign indications, black women were less likely to receive minimally invasive hysterectomy compared to white women. Future studies are needed to further explore the factors responsible for this possible racial disparity in hysterectomy options, which may include patients’ medical complexity, size of uterine fibroids, and limited training for surgeons to perform MIH in the presence of uterine fibroids.
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Patients: A systematic search of literature on all material available on methods to diagnose AUB was performed from January 2001 till April 2017. Intervention: nil. Measurements and Main Results: In majority of the studies it has been proven that saline infusion sonography (SIS) is definitely superior to plain TVS and especially helpful in differentiating focal and global pathologies. Although hysteroscopy guided biopsy is considered the gold standard, sonohysterography acts as a useful guide. SIS and Hysteroscopy were compared for the following parameters: sensitivity, specificity, positive predictive value, negative predictive value & kappa value. Conclusion: SIS serves as an “indispensable tool” which has successfully created its own space between Trans Vaginal Sonography. Growing expertise in the field of SIS may reduce the number of hysteroscopy. The introduction of gel instead of saline and 3D technology is promising. SIS directed biopsies may even replace the need for Hysteroscopy as a diagnostic tool. 319
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:51 AM – STATION A
SLC40A1 Sensitized Human Ovarian Cancer Cells to Cisplatin by Blocking Autophagy Bao L, Wu J, Yi X. Department of Gynecology, Obstetrics and Gynecology Hospital Fudan University, Shanghai, China Study Objective: This study aimed to investigate the role of the iron export related gene, solute carrier family 40 member 1 (SLC40A1), in cisplatin resistance of epithelial ovarian cancer (EOC) and to explore the underlying mechanism. Design: In vitro study. Setting: A university-based tertiary obstetrics and gynecology hospital. Patients: N/A. Intervention: N/A. Measurements and Main Results: Expression of SLC40A1 in normal epithelial ovarian tissue and EOC tissue was examined by western blot and realtime PCR. Two pairs of cisplatin-sensitive (A2780 and COC1) and cisplatin-resistant (A2780cp and COC1/DDP) ovarian cancer cells were used to detect the expression of SLC40A1. SLC40A1 was overexpressed in normal epithelial ovarian tissue, while relatively lower in low-grade serous ovarian cancer, and the lowest in high-grade serous ovarian cancer.
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:45 AM – STATION H
Saline Infusion Sonography an “Indispensable Tool” or “Superfluous Trinket” in Diagnosis of Abnormal Uterine Bleeding in Comparison with Hysteroscopy Agrawal S, Goenka S. Obstetrics & Gynecology, All India Institute of Medical Sciences, Raipur, Chhattisgarh, India Study Objective: To review and summarize the current evidence-based literature on the role of saline infusion sonography in comparison to other available diagnostic methods in cases of abnormal uterine bleeding. Design: : A systematic search of literature on all material available on methods to diagnose AUB was performed from January 2001 till April 2017 using key words like abnormal uterine bleeding, saline Infusion sonography, sonohysterography, TVS in AUB, Hysteroscopy in AUB. Articles were included after assessing for quality and suitability for inclusion in the following order like systemic reviews, Meta analysis, guidelines, RCT, prospective cohort studies, observational studies, non systemic reviews and case series. Setting: Department of Obstetrics & Gynecology, All India Institute of Medical Sciences Raipur Chhattisgarh.
Both chemo-sensitive ovarian cancer cell line A2780 and COC1 showed relatively higher level of SLC40A1 and lower intracellular iron compared with their corresponding chemo-resistant counterparts. Along with knockdown of SLC40A1 in A2780 and COC1, the intracellular iron increased, while cell viability and apoptosis rate decreased significantly. Along with SLC40A1
Abstracts / Journal of Minimally Invasive Gynecology 24 (2017) S1–S201 overexpression in A2780cp and COC1/DDP, the intracellular iron decreased, while cell viability and apoptosis rate increased significantly. Further, we examined the autophagy gene expression both in SLC40A1 knockdown and upregulated cells. Interestingly, autophagy-related gene Beclin 1 and Atg3 increased along with SLC40A1 knockdown and decreased along with SLC40A1 upregulated both at mRNA and protein level. Moreover, the transmission electron microscopy examination showed that the autophagosome was blocked along with SLC40A1 overexpression.
Conclusion: This is the first study suggesting that the SLC40A1-mediated iron metabolism contribute to cisplatin resistance through blocking autophagy in EOC.
320
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:51 AM – STATION B
Successive Fertility Following Optimized Perfusion and Cryopreservation of Whole Ovary and Allotransplantation in a Premature Ovarian Insufficiency Rat Model Ding Y,1 Shao J,2 Li J,1 Hong K,1 Zhang Y,1 Hua K,1 Wang X2. 1 Department of Gynecology, Obstetrics and Gynecology Hospital, Fudan
University, Shanghai, China; 2Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China Study Objective: To optimize perfusion, and to explore the possibility of restoring ovarian function and natural fertility after whole ovarian cryopreservation and transplantation (WOCP&TP) to a premature ovarian insufficiency (POI) rat model. The effects of cryopreservation on offsprings of rats following WOCP&TP were also investigated. Design: The ovarian function and fertility were observed after WOCP&TP with optimized perfusion in 25 CTX-induced POI rats for 8 months. Setting: Animal Experiment Efficacy Evaluation Center, School of Pharmacy, Fudan University, Shanghai, China. Patients: Lewis rats aged 8–10 weeks weighing 180-200g. Intervention: The whole ovaries were observed in morphology and TUNEL assay after optimized perfusion. The ovarian function was assessed by vaginal smears and blood hormonal levels. Fertility restoration was quantified by live birth rates after mating and appearance of offsprings. Measurements and Main Results: Histological evaluation demonstrated intact morphology of follicles in all groups, with 77.6% of the total follicle in optimized group. The apoptosis rates of ovarian cells in optimized group were significantly reduced compared to the conventional group.
Fig 1. The apoptosis rates of ovarian cells in different groups.Compared to control group, *P<0.05, **P<0.01; compared to conventional group, ## P<0.01.
Of the 25 recipient POI rats, 5 rats were died. Fourteen rats began to recover ovarian function after 2 weeks of transplantation, with normal hormone levels after 4 weeks of transplantation. Four in 14 rats were pregnant and delivered live births. One rat was given second time pregnancy and delivered live birth. The first generation of young rats gave birth of the second generation of rats. After mating, the second generation of young rats gave birth of the third generation of rats. No abnormalities were found in all the offsprings. Conclusion: High rates of restoration of ovarian function and live birth rates were obtained following WOCP&TP to POI rat models utilizing optimized perfusion. Cryopreservation could not affect the nature of successive generations.
Follicles in representative sections of ovaries in different groups Groups Number of follicles primordial
primary
secondary
antral
Optimized group (n=5) Conventional group (n=5) Control group (n=5)
22.5±2.1* 17.0±1.8** 28.3±3.5
9.8±1.0* 7.5±1.3** 12.5±1.7
3.8±1.0** 3.0±0.8** 7.0±0.8
79.8±7.0** 66.3±5.9** 104.8±6.7
Compared to control group, *P<0.05, **P<0.01.
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total number of follicles 118.0±6.9** (77.6%) 94.5±7.3** (61.0%) 152.5±11.6 (100%)
Abstracts / Journal of Minimally Invasive Gynecology 24 (2017) S1–S201 Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:45 AM – STATION G
Influence of Race/Ethnicity on Route of Hysterectomy and Inpatient Surgical Complications Bougie O,1 Singh SS,1 McCarthy EP2. 1Ottawa Hospital Research Institute, University of Ottawa, Ottawa, Ontario, Canada; 2Harvard T.H. Chan School of Public Health, Boston, Massachusetts Study Objective: To examine the association between race and (1) route of hysterectomy and (2) risk of inpatient surgical complications. Design: A cross-sectional analysis of women who underwent hysterectomy during 2009–2013 using the Nationwide Inpatient Sample. Setting: The Nationwide Inpatient Sample (NIS) is 20% stratified sample of all patient discharges from acute care hospitals in the United States designed to be nationally representative. Patients: We identifed 114,719 women age 18 and older who underwent elective hysterectomy for benign indications using International Classification of Diseases (ICD-9-CM) procedure and diagnostic codes. Intervention: Our main factor of interest was race/ethnicity, categorized as: White, Black, Hispanic, Other or missing. Measurements and Main Results: Compared to white women, black women had 0.55 (95%CI: 0.52–0.59) odds of receiving minimally invasive hysterectomy, adjusted for age, Median household income national quartile for patient ZIP Code, primary payer, hospital location/teaching status, Elixhauser comorbidity index, and indication for surgery. This finding remained consistent across household income quartile for patient ZIP Code, primary payer and diagnosis of fibroids. In our sample of women, 6091 experienced a complication, representing 30,455 women nationwide. Compared to white women, the odds of black women experiencing a surgical complication was 1.03(95% CI: 0.93– 1.13) after adjusting for age, median household income national quartile for patient ZIP Code, primary payer, hospital location/teaching status, Elixhauser comorbidity index, and route of surgery. This finding remained consistent across household income quartile for patient ZIP Code, primary payer and route of hysterectomy. Conclusion: Among women who underwent hysterectomy for benign indications, black women were less likely to receive minimally invasive hysterectomy compared to white women. Future studies are needed to further explore the factors responsible for this possible racial disparity in hysterectomy options, which may include patients’ medical complexity, size of uterine fibroids, and limited training for surgeons to perform MIH in the presence of uterine fibroids.
318
Patients: A systematic search of literature on all material available on methods to diagnose AUB was performed from January 2001 till April 2017. Intervention: nil. Measurements and Main Results: In majority of the studies it has been proven that saline infusion sonography (SIS) is definitely superior to plain TVS and especially helpful in differentiating focal and global pathologies. Although hysteroscopy guided biopsy is considered the gold standard, sonohysterography acts as a useful guide. SIS and Hysteroscopy were compared for the following parameters: sensitivity, specificity, positive predictive value, negative predictive value & kappa value. Conclusion: SIS serves as an “indispensable tool” which has successfully created its own space between Trans Vaginal Sonography. Growing expertise in the field of SIS may reduce the number of hysteroscopy. The introduction of gel instead of saline and 3D technology is promising. SIS directed biopsies may even replace the need for Hysteroscopy as a diagnostic tool. 319
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:51 AM – STATION A
SLC40A1 Sensitized Human Ovarian Cancer Cells to Cisplatin by Blocking Autophagy Bao L, Wu J, Yi X. Department of Gynecology, Obstetrics and Gynecology Hospital Fudan University, Shanghai, China Study Objective: This study aimed to investigate the role of the iron export related gene, solute carrier family 40 member 1 (SLC40A1), in cisplatin resistance of epithelial ovarian cancer (EOC) and to explore the underlying mechanism. Design: In vitro study. Setting: A university-based tertiary obstetrics and gynecology hospital. Patients: N/A. Intervention: N/A. Measurements and Main Results: Expression of SLC40A1 in normal epithelial ovarian tissue and EOC tissue was examined by western blot and realtime PCR. Two pairs of cisplatin-sensitive (A2780 and COC1) and cisplatin-resistant (A2780cp and COC1/DDP) ovarian cancer cells were used to detect the expression of SLC40A1. SLC40A1 was overexpressed in normal epithelial ovarian tissue, while relatively lower in low-grade serous ovarian cancer, and the lowest in high-grade serous ovarian cancer.
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:45 AM – STATION H
Saline Infusion Sonography an “Indispensable Tool” or “Superfluous Trinket” in Diagnosis of Abnormal Uterine Bleeding in Comparison with Hysteroscopy Agrawal S, Goenka S. Obstetrics & Gynecology, All India Institute of Medical Sciences, Raipur, Chhattisgarh, India Study Objective: To review and summarize the current evidence-based literature on the role of saline infusion sonography in comparison to other available diagnostic methods in cases of abnormal uterine bleeding. Design: : A systematic search of literature on all material available on methods to diagnose AUB was performed from January 2001 till April 2017 using key words like abnormal uterine bleeding, saline Infusion sonography, sonohysterography, TVS in AUB, Hysteroscopy in AUB. Articles were included after assessing for quality and suitability for inclusion in the following order like systemic reviews, Meta analysis, guidelines, RCT, prospective cohort studies, observational studies, non systemic reviews and case series. Setting: Department of Obstetrics & Gynecology, All India Institute of Medical Sciences Raipur Chhattisgarh.
Both chemo-sensitive ovarian cancer cell line A2780 and COC1 showed relatively higher level of SLC40A1 and lower intracellular iron compared with their corresponding chemo-resistant counterparts. Along with knockdown of SLC40A1 in A2780 and COC1, the intracellular iron increased, while cell viability and apoptosis rate decreased significantly. Along with SLC40A1
Abstracts / Journal of Minimally Invasive Gynecology 24 (2017) S1–S201 overexpression in A2780cp and COC1/DDP, the intracellular iron decreased, while cell viability and apoptosis rate increased significantly. Further, we examined the autophagy gene expression both in SLC40A1 knockdown and upregulated cells. Interestingly, autophagy-related gene Beclin 1 and Atg3 increased along with SLC40A1 knockdown and decreased along with SLC40A1 upregulated both at mRNA and protein level. Moreover, the transmission electron microscopy examination showed that the autophagosome was blocked along with SLC40A1 overexpression.
Conclusion: This is the first study suggesting that the SLC40A1-mediated iron metabolism contribute to cisplatin resistance through blocking autophagy in EOC.
320
Virtual Posters – Session 1 (9:45 AM - 10:45 AM) 9:51 AM – STATION B
Successive Fertility Following Optimized Perfusion and Cryopreservation of Whole Ovary and Allotransplantation in a Premature Ovarian Insufficiency Rat Model Ding Y,1 Shao J,2 Li J,1 Hong K,1 Zhang Y,1 Hua K,1 Wang X2. 1 Department of Gynecology, Obstetrics and Gynecology Hospital, Fudan
University, Shanghai, China; 2Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China Study Objective: To optimize perfusion, and to explore the possibility of restoring ovarian function and natural fertility after whole ovarian cryopreservation and transplantation (WOCP&TP) to a premature ovarian insufficiency (POI) rat model. The effects of cryopreservation on offsprings of rats following WOCP&TP were also investigated. Design: The ovarian function and fertility were observed after WOCP&TP with optimized perfusion in 25 CTX-induced POI rats for 8 months. Setting: Animal Experiment Efficacy Evaluation Center, School of Pharmacy, Fudan University, Shanghai, China. Patients: Lewis rats aged 8–10 weeks weighing 180-200g. Intervention: The whole ovaries were observed in morphology and TUNEL assay after optimized perfusion. The ovarian function was assessed by vaginal smears and blood hormonal levels. Fertility restoration was quantified by live birth rates after mating and appearance of offsprings. Measurements and Main Results: Histological evaluation demonstrated intact morphology of follicles in all groups, with 77.6% of the total follicle in optimized group. The apoptosis rates of ovarian cells in optimized group were significantly reduced compared to the conventional group.
Fig 1. The apoptosis rates of ovarian cells in different groups.Compared to control group, *P<0.05, **P<0.01; compared to conventional group, ## P<0.01.
Of the 25 recipient POI rats, 5 rats were died. Fourteen rats began to recover ovarian function after 2 weeks of transplantation, with normal hormone levels after 4 weeks of transplantation. Four in 14 rats were pregnant and delivered live births. One rat was given second time pregnancy and delivered live birth. The first generation of young rats gave birth of the second generation of rats. After mating, the second generation of young rats gave birth of the third generation of rats. No abnormalities were found in all the offsprings. Conclusion: High rates of restoration of ovarian function and live birth rates were obtained following WOCP&TP to POI rat models utilizing optimized perfusion. Cryopreservation could not affect the nature of successive generations.
Follicles in representative sections of ovaries in different groups Groups Number of follicles primordial
primary
secondary
antral
Optimized group (n=5) Conventional group (n=5) Control group (n=5)
22.5±2.1* 17.0±1.8** 28.3±3.5
9.8±1.0* 7.5±1.3** 12.5±1.7
3.8±1.0** 3.0±0.8** 7.0±0.8
79.8±7.0** 66.3±5.9** 104.8±6.7
Compared to control group, *P<0.05, **P<0.01.
S107
total number of follicles 118.0±6.9** (77.6%) 94.5±7.3** (61.0%) 152.5±11.6 (100%)