Metformin enhances epithelial cell growth inhibition via the protein kinase-insulin-like growth factor binding protein-1 pathway.

BACKGROUND: Abnormal stromal-epithelial cell communication is a pathogenic mechanism in endometriosis, and metformin can modulate it. Insulin-like growth factor binding protein-1 (IGFBP1) plays a role in endometriosis, but the exact mechanism is unknown. IGFBP1 is reportedly a downstream target of metformin in some diseases. We aimed to investigate the role of IGFBP1 in endometriosis development, whether it is associated with abnormal communication, and whether metformin affects IGFBP1 expression. METHODS: Patients who underwent surgical treatment for endometriosis or other diseases were enrolled. Ten patients with ovarian-type endometriosis and eight patients each who underwent surgical treatment for other lesions with or without endometriosis were selected, and their tissues taken for cell proliferation, western blotting, polymerase chain reaction, and knockdown experiments. RESULTS: Ectopic and eutopic stromal cells (EcSCs and EuSCs) lost their ability to inhibit epithelial cell proliferation, and IGFBP1 expression was downregulated in both groups of stromal cells compared to that in normal stromal cells (NSCs; 1.09 vs. 0.25, p = .0002 1.09 vs. 0.57, p = .0029). In an EcSC IGFBP1 overexpression model, the ability of EcSCs to inhibit epithelial cell proliferation was enhanced (EdU positivity decreased from 38% to 25%, p = .0001). Furthermore, adenosine 5'-monophosphate-activated protein kinase (AMPK) phosphorylation was downregulated in EcSCs and EuSCs compared to that in NSCs (0.99 vs. 0.42, p = .0006/0.99 vs. 0.57, p = 0.0032). Treatment of EcSCs with metformin increased AMPK phosphorylation (0.47 vs. 1.04, p = .0107) while upregulating IGFBP1 expression (0.69 vs. 1.01, p = .0164), whereas pre-treatment with an AMPK phosphorylation inhibitor abrogated metformin-induced IGFBP1 upregulation. CONCLUSIONS: IGFBP1 mediates aberrant stromal-epithelial communication in endometriosis. Metformin can upregulate IGFBP1 expression in EcSCs by activating AMPK, and upregulated IGFBP1 enhances the inhibition of epithelial cell proliferation. IGFBP1 is expected to be a therapeutic target for endometriosis.

Insulin-like growth factor binding protein 1 (IGFBP1) is a protein that regulates cell growth and proliferation and is expressed at abnormal levels in patients with endometriosis. In some cases, metformin has been shown to modulate the expression of this protein. Here, we investigated the role of IGFBP1 in endometriosis development, whether it is associated with abnormal communication, and whether metformin affects IGFBP1 expression in endometrial cells. We found that downregulation of IGFBP1 in endometriosis diminished the ability of stromal cells to inhibit the proliferation of epithelial cells through inhibition of the protein kinase B and extracellular regulated protein kinase pathways. In addition, metformin upregulated IGFBP1 expression by activating adenosine 5'-monophosphate-activated protein kinase, suggesting that IGFBP1 may be one of the potential targets for drug therapy for endometriosis.

Impact on ovarian reserve function by different homostasis methods during laparoscopic cystectomy in treatment of ovarian endometrioma / 中华妇产科杂志

Objective To investigate the impact on ovarian reserve function by different hemostasis methods during laparoscopic surgery in treatment of ovarian endometrioma.Methods From September 2008 to February 2010,162 cases with ovarian endometrioma undergoing laparoscopic surgery in Shandong Provincial Hospital were enrolled in this study.At the 3rd day of the menstrual cycle before surgery and the 1 st,3rd,6th and 12th cycle after surgery,serum FSH and anti-mullerian hormone(AMH) and ultrasound basal antral follicle count (AFC) and peak systolic velocity (PSV) were examined and compared.Based on hemostasis method,those patients were divided into 3 groups,including 54 cases in bipolar hemostasis,54 cases in ultrasonic scalpel hemostasis and suture after excision of endometrioma.Results (1) Before surgery:no significant different factors among three groups before surgery were observed,including age,size of endometrioma,the level of FSH,AMH,AFC,PSV (P ＞ 0.05).(2) Ovarian reserve function after surgery:①FSH:at the 1st,3rd,6th and 12th month follow-up,the FSH in the bipolar group was (11.7 ±4.0),(9.9 ± 4.0),(9.5 ± 4.3),(9.5 ± 3.9) U/L,and the FSH in ultrasonic scalpel group was (11.4 ±4.3),(9.7 ± 4.0),(9.2 ± 3.7),(9.9 ± 4.6) U/L,were significantly higher than (9.3 ± 3.8),(6.7 ±3.0),(6.5 ± 3.2),(6.4 ± 2.2) U/L in suture group respectively (all P ＜ 0.05).()AMH:at the 1 st,3rd,6th and 12th month follow-up,the AMH in the bipolar group was (1.8 ±0.9),(1.8 ± 1.0),(1.9 ±1.0),(2.0 ± 1.0) μg/L,and the AMH in the ultrasonic scalpel group was (1.6 ±± 0.8),(1.8 ± 1.0),(2.0 ± 1.1),(2.1 ± 1.0) μg/L,which were significantly lower than (2.8 ± 1.7),(2.9 ± 1.6),(3.0 ±1.3),(3.2 ± 1.5) μg/L in suture group,respectively (all P ＜ 0.05).③AFC:there was no significant difference of APC among the three groups in the 1st month after surgery.However,at the 3rd,6th and 12th month follow-up,the AFC of 4.8 ± 1.4,5.9 ± 1.5,6.1 ± 1.5 in the suture group was significant higher than 3.7 ± 1.4,4.1 ± 1.4,4.0 ± 1.5 in bipolar group and 3.6 ± 1.3,4.0 ± 1.1,3.9 ± 1.5 in ultrasonic group,respectively (all P ＜ 0.05).④PSV:at the 1 st,3rd,6th and 12th month follow-up,the PSV of the bipolar group(7.9 ±3.5),(8.1 ±3.3),(8.4 ±3.1),(8.6±3.0) cm/s in bipolar group and (8.1 ±3.5),(8.0 ± 3.0),(7.9 ± 3.2),(8.0 ± 2.9) cm/s in ultrasonic group were significant lower than (10.9 ± 3.3),(12.0 ± 3.2),(11.8 ± 3.0),(12.1 ± 4.1) cm/s in suture group,respectively.(allP＜0.05).Conclusions Bipolar or ultrasonic scalpel hemostasis during laparoscopic excision of ovarian endometrioma is associated with a significant reduction in ovarian reserve.Electrocoagulation of the ovarian tissue should be avoided.

Study on the treatment of high dose mifepristone and progesterone in endometrial carcinoma / 中华妇产科杂志

Objective To investigate the effect of high dose mifepristone and high dose progesterone in the treatment of patients with endometrial carcinoma and to explore the possible mechanisms associating with them Methods Thirty untreated patients diagnosed as endometrial carcinoma through dilation and curettage of the uteri were divided into 3 groups at random Each group was given medroxyprogesterone acetate(MPA),(500 mg/day) or mifepristone(MIF),(100 mg/day)or MIF(100 mg/day)+ MPA(500 mg/day)for 5 days respectively On the sixth day, hysterectomy was performed on these patients The endometrial cancer specimen of post hysterectomy was compared with the one of pre administrating The morphologic changes of the endometrial cancer cells were observed through light microscope Immunohistochemistry assay (SP method) was applied to determine the localization and immunoreactive intensity of proliferating cell nuclear antigen(PCNA), estrogen receptor (ER), progesterone receptor (PR), B cell leukemia lymphoma 2 (bcl 2), bcl 2 associated X protein(bax) and CD 44 v6 Results Better differentiation degree and active excretion were observed in all of the post hysterectomy endometrial specimen In the same time, apoptosis of carcinoma cells was observed The most significant changes were seen in the MIF+MPA group In the MPA group,the pre treatment and post treatment expression of PR(2 9?1 1,1 6?0 8),ER(2 8?0 9,1 4?0 9),PCNA(0 84?0 10,0 60?0 12),bcl 2(0 236?0 089,0 157?0 981) and CD 44 v6 (4 6?1 8,2 5?1 9) were all decreased(all P 0 05) In the MIF+MPA group, the expression of PR(3 2?1 0,0 8?0 8),ER(2 7?0 9,0 7 ?0 9 ),PCNA(0 81?0 09,0 25?0 09),bcl 2(0 225?0 091,0 066?0 009)and CD 44 v6(4 5?1 9,2 7?1 6) were all decreased(all P

Effect of fetal liver AFT024 cells on multidrug resistant gene 1 transfection efficiency and in vitro expansion of CD34~+ cells derived from umbilical cord blood / 中国肿瘤生物治疗杂志

Objective:To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1(MDR1)and the in vitro expansion of CD34+ cells derived from umbilical cord blood.Methods:CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and co-cultured with AFT024 cells(AFT024 group)or cultured alone(control group)for 7 days.During the subsequent 14 days,retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells.On the 7th,14th and 21st day after culture,the number of total nucleated cells(TNC)was counted,the ratio of CD34+ cells was assayed by flow cytometry(FCM)and the number of CD34+ cells was calculated,and colony-forming cells(CFC)were counted by methylcellulose cultures.RT-PCR method was used to detect the level of MDR1 mRNA in the transfected cells.The expression and function of P-glycoprotein(P-gp)were evaluated by FCM assay and Rhodamine-123 efflux assay,respectively.The gene transfection efficiency was calculated by drug-resistant colony-forming cells assay.Results:(1)The MDR1 mRNA level in AFT024 group than that in control group.The gene transfection efficiency in AFT024 group was significantly higher than that in control group(46.0% vs 15.2%,P0.05).On the 14th day,the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P0.05).The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P