Genistein attenuates isoflurane-induced neurotoxicity and improves impaired spatial learning and memory by regulating cAMP/CREB and BDNF-TrkB-PI3K/Akt signaling.

Anesthetics are used extensively in surgeries and related procedures to prevent pain. However, there is some concern regarding neuronal degeneration and cognitive deficits arising from regular anesthetic exposure. Recent studies have indicated that brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB) are involved in learning and memory processes. Genistein, a plant-derived isoflavone, has been shown to exhibit neuroprotective effects. The present study was performed to examine the protective effect of genistein against isoflurane-induced neurotoxicity in rats. Neonatal rats were exposed to isoflurane (0.75%, 6 hours) on postnatal day 7 (P7). Separate groups of rat pups were orally administered genistein at doses of 20, 40, or 80 mg/kg body weight from P3 to P15 and then exposed to isoflurane anesthesia on P7. Neuronal apoptosis was detected by TUNEL assay and FluoroJade B staining following isoflurane exposure. Genistein significantly reduced apoptosis in the hippocampus, reduced the expression of proapoptotic factors (Bad, Bax, and cleaved caspase-3), and increased the expression of Bcl-2 and Bcl-xL. RT-PCR analysis revealed enhanced BDNF and TrkB mRNA levels. Genistein effectively upregulated cAMP levels and phosphorylation of CREB and TrkB, leading to activation of cAMP/CREB-BDNF-TrkB signaling. PI3K/Akt signaling was also significantly activated. Genistein administration improved general behavior and enhanced learning and memory in the rats. These observations suggest that genistein exerts neuroprotective effects by suppressing isoflurane-induced neuronal apoptosis and by activating cAMP/CREB-BDNF-TrkB-PI3/Akt signaling.

The Treatment of Pelvic Locoregional Recurrence of Cervical Cancer After Radical Surgery With Intensity-Modulated Radiation Therapy Compared With Conventional Radiotherapy: A Retrospective Study.

OBJECTIVE: The aim of this study was to investigate the therapeutic response and toxicity of intensity-modulated radiation therapy (IMRT) or conventional radiotherapy (c-RT) as adjuvant therapy in patients with pelvic locoregional recurrence of cervical cancer after radical surgery. METHODS: This retrospective study included 161 patients with unresectable pelvic locoregional recurrence of cervical cancer after radical surgery between March 2003 and May 2012. All patients were initially diagnosed with International Federation of Gynecology and Obstetrics stage IB-IIA cervical cancer and received radical hysterectomy and pelvic lymphadenectomy. A total of 82 patients were treated with c-RT, whereas the remaining 79 patients underwent IMRT. Intracavitary brachytherapy and concurrent chemotherapy were performed during external irradiation. RESULTS: The mean dose delivered to the planning target volume was significantly higher in the IMRT group than in the c-RT group (61.8 vs 50.3 Gy, P = 0.029). Intensity-modulated radiation therapy plans yielded better dose sparing of small bowel, bladder, and rectum than did c-RT (P < 0.05). Moreover, the IMRT patients experienced less acute and chronic toxicities (P < 0.05) and better short-term effects (complete response + partial response) than did those treated with c-RT (89.9% vs 63.4%, P = 0.03). Three- and 5-year overall survival rates were significantly higher in the IMRT group than in the c-RT group (3-year: 58.4% vs 39.1%, P = 0.012; 5-year: 35.4% vs 21.4%, P = 0.007). Furthermore, 5-year progression-free survival rates were significantly higher in the IMRT group than in the c-RT group (26.1% vs 15.1%, P = 0.031). CONCLUSIONS: Intensity-modulated radiation therapy achieved outcomes superior to c-RT in patients with pelvic locoregional recurrence of cervical cancer after radical surgery. The acute and chronic toxicities were acceptable, and the adjacent organs at risk were well protected.

Prognostic significance of reduced immunohistochemical expression of E-cadherin in endometrial cancer-results of a meta-analysis.

OBJECTIVE: Previous studies which investigated the relationship between reduced E-cadherin and prognosis of endometrial cancer were ambiguous and conflicting. Therefore, the aim of the present study was to evaluate the relationship between reduced expression of E-cadherin and endometrial cancer by meta-analysis approach. METHOD: AfterPubmed and Embasewere deliberately searched via the internet, 8 pieces of literaturewere totally included in final meta-analysis. After the data had been abstracted, the pulled odds ratio (OR) and hazard ratio (HR) were calculated by STATA with random or fixed effect model depending on their heterogeneity. The publication bias of included literature were tested by Begg's funnel plot and Egger's test. RESULTS: The pulled data showed that the reduced expression of E-cadherin was significantly associated with overall survival (OS), HR=2.42, 95% CI: 1.50-3.89. The clinical parameters such as lymph node metastasis (LNM), myometrial invasion (MI), International Federation of Gynecology and Obstetrics (FIGO) stage, histological type and pathological type were also significantly associated with reduced expression of E-cadherin. The results of publication biasshowed there were no significant publication bias. CONCLUSION: Endometrial cancer patients with reduced expression of E-cadherin may have a poorer prognosis than those with normal or higher expression of E-cadherin.

[Effects of different soil types on the foliar δ13C values of common local plant species in karst rocky desertification area in central Guizhou Province].

By measuring the foliar δ13C values of common local plant species grown in different soil types in Wangjiazhai catchments, a typical karst desertification area in Qingzhen City, Central Guizhou, we studied the impact of soil type and rocky desertification grade on the foliar δ13C values. The results showed that the foliar δ13C values were more negative in yellow soil area than those in black calcareous area and there was no obvious difference in foliar δ13C values between these two soil types. The distribution interval of foliar δ13C values in yellow soil area was narrower than those in black calcareous area and the variation coefficient of foliar δ13C values in yellow soil area were smaller than those in black calcareous area. With increasing degree of karst rocky desertification, the foliar δ13C values of plant community in black calcareous area increased, whereas those in yellow soil area first increased and then decreased. The result of multiple comparison showed that the difference in foliar δ13C values of plant community among rocky desertification grade was not obvious in yellow soil area, but it was obvious in black calcareous area. Correlation analysis between the foliar δ13C values of plant species and the main environmental factors indicated that slope and soil thickness were the main factors which affected the foliar δ13C values of plants in yellow soil area and soil water contant was the main factor in black calcareous area. The impact of soil on the foliar δ13C values was realized by adjusting the soil moisture in study area.

[Clinical study of sentinel lymph node detection guided radical abdominal trachelectomy].

OBJECTIVE: To evaluate the clinical value of sentinel lymph nodes (SLN) in predicting pelvic lymph node status for early cervical squamous cell carcinoma, and approach the clinical significance of SLN detection for guiding radical abdominal trachelectomy (RAT).Outcomes of follow up and fertility were also observed. METHODS: A total of 31 patients with stageIa2-Ib1 squamous cell carcinoma planned to be given RAT and pelvic lymphadenectomy were enrolled. (99m)Tc-labeled phytate was injected before surgery.Intraoperatively, SLN were identified, excised, and submitted to fast frozen section.Systematic bilateral pelvic lymphadenectomy was performed, and then RAT was performed in patients with negative SLN. All nodes were sent for routine pathological examination and immunostained with anti-cytokeratin antibody to detect micrometastases. RESULTS: SLN were detected in all patients (100%,31/31). A total of 109 SLN were identified with a mean number of 3.5 per patient.Of these, SLN of 2 patients were positive on frozen sections and proved to be metastasis by final pathologic examination and quitted the RAT. No missed micrometastasis was found using immunohistochemical staining in SLN and other lymph nodes using histologically node-negative cases. No false negative cases was found and the negative value was 100% (31/31). The sensitivity, accuracy, and false negative rates were 100%, 100%, and 0, respectively. Perioperative complications occurred in 5 patients including 2 cases of bladder injury and 3 cases of uterine artery injury.No relapses occurred during follow-up.Five of 19 patients with procreative desire conceived pregnancies (4 spontaneous abortion and 1 premature birth) after surgery. CONCLUSIONS: The identification of SLN using (99m)Tc-labeled phytate could predict the pelvic lymph node status in early stage cervical cancer. Under the guidance of SLN detection, RAT is a feasible operative modality with well prognosis and low complications for young patients who desire to preserve reproductive function.

[Application value of reduced field intensity modulated radiation therapy for advanced cervical cancer].

OBJECTIVE: To explore the clinical value and efficacy of reduced field intensity modulated radiation therapy (RF-IMRT) for patients with advanced cervical cancer. METHODS: Seventy-one patients with stage IIB-IIIB cervical cancer, who underwent reduced field IMRT (RF-IMRT group) and 72 patients treated with conventional radiotherapy (c-RT group) in Shandong Cancer Hospital between 2005 August and 2011 August, were enrolled in this study. The RF-IMRT plans were as follows: whole pelvic IMRT plan was performed to deliver an initial dose of 30 Gy, then the irradiated volume was reduced to lymphatic drainage region as well as paracervix and parametrium for an additional 30 Gy boost. Conventional 2-field RT plan was performed in these patients using ADAC Pinnacle 3 planning system, to be given the same prescription dose, and to compare the irradiation dose of organs at risk (OARs). At the same time, conventional 2-field RT was performed in 72 patients of the c-RT group. Concurrent chemotherapy and intracavitary brachytherapy were also performed in the two groups. The treatment response, toxicities, normal tissue avoidance, and survival were assessed. RESULTS: Sixty-six patients of the RF-IMRT group and 65 patients of the c-RT group fulfilled the treatment plan. IMRT plans yielded better dose conformity to the target (0.711 ± 0.057 vs. 0.525 ± 0.062, P = 0.032) and better sparing of the rectum, bladder and small intestine (rectum: 41.6 ± 6.8 vs. 50.8 ± 3.2, P = 0.016; bladder: 40.2 ± 2.9 vs. 51.4 ± 1.8, P = 0.007; small intestine: 22.3 ± 2.6 vs. 35.8 ± 3.9, P = 0.004). The mean dose delivered to the planning target volume (PTV) was significantly higher in the RF-IMRT group than that in the c-RT group (60.8 vs. 51.2 Gy, P = 0.006). The RF-IMRT patients experienced significantly lower acute and chronic toxicities with comparable short-term effects than did those treated with conventional RT (P > 0.05). No significant differences were found between the two groups for 1-, 3-, and 5-year overall survival (OS) rates, while a significantly higher progression-free survival (PFS, 65.2% vs. 46.2%, P = 0.031) rate was observed in the RF-IMRT group. CONCLUSIONS: RF-IMRT yields higher dose distributions and lower toxicities compared with conventional RT, and both the tumor target volume and pelvic lymphatic drainage region achieve curative dose irradiation, the adjacent organs at risk are well protected, and with tolerable adverse reactions. Yet, RF-IMRT provides comparable clinical outcomes and higher PFS.

[Isolation, purification and characterization of endothelial cells from endometrial cancer and normal endometrial tissue].

OBJECTIVE: To immunopurify human endometrial endothelial cells (HEEC) from fresh surgical specimens of endometrial cancers and normal endometrial tissues, and investigate their biological characteristics. METHODS: Endothelial cells of endometrial cancers and normal endometrial tissues were isolated using anti-CD31 conjugated magnetic microbeads. The isolated endothelial cells were cultured in vitro and their origins were identified. Their angiogenic characteristics were observed by MTT, wound healing, Transwell cell invasion and tube formation assays. RESULTS: Flow cytometry revealed that the immunopurification technique yielded endothelial cell purity of > 95% in all samples. All purified HEEC were characterized as endothelial cells on the basis of expression of the classical endothelial markers vWF and CD31 as shown by immunofluorescence examination. Although the tumor-associated HEEC didn't show more rapid proliferation than normal HEEC, they exhibited enhanced migration ability (P = 0.006), potent invasiveness (P = 0.033), and elevated tube formation in vitro (P = 0.029). CONCLUSIONS: Human endometrial endothelial cells can be efficiently isolated from endometrial cancer and normal endometrial tissues by immunomagnetic methods. Tumor-associated HEEC exhibit enhanced migratory ability, potent invasiveness, and elevated tube formation in vitro.

PET/CT scanning guided intensity-modulated radiotherapy in treatment of recurrent ovarian cancer.

OBJECTIVE: This study was undertaken to evaluate the clinical contribution of positron emission tomography using (18)F-fluorodeoxyglucose and integrated computer tomography (FDG-PET/CT) guided intensity-modulated radiotherapy (IMRT) for treatment of recurrent ovarian cancer. MATERIALS AND METHODS: Fifty-eight patients with recurrent ovarian cancer from 2003 to 2008 were retrospectively studied. In these patients, 28 received PET/CT guided IMRT (PET/CT-IMRT group), and 30 received CT guided IMRT (CT-IMRT group). Treatment plans, tumor response, toxicities and survival were evaluated. RESULTS: Changes in GTV delineation were found in 10 (35.7%) patients based on PET-CT information compared with CT data, due to the incorporation of additional lymph node metastases and extension of the metastasis tumor. PET/CT guided IMRT improved tumor response compared to CT-IMRT group (CR: 64.3% vs. 46.7%, P=0.021; PR: 25.0% vs. 13.3%, P=0.036). The 3-year overall survival was significantly higher in the PET-CT/IMRT group than control (34.1% vs. 13.2%, P=0.014). CONCLUSIONS: PET/CT guided IMRT in recurrent ovarian cancer patients improved the delineation of GTV and reduce the likelihood of geographic misses and therefore improve the clinical outcome.

[Angiogenesis related gene expression profiles of human endometrial endothelial cells isolated from tissue of endometrial cancer].

OBJECTIVE: To identify and verify the different genes expression pattern between human endometrial endothelial cells (HEEC) isolated from endometrial cancer and normal endometrium. METHODS: Endothelial cells were isolated from 5 patients with endometrial cancer (endometrial cancer group 1) and 5 patients with normal endometria tissue (control group 1) admitted from June to November 2007 in Shandong Cancer Hospital. Global expression patterns of endothelial cells were examined using oligonucleotide microarrays. Tissues from 36 patients with endometrial cancer (endometrial cancer group 2) and 10 normal endometrial tissues (control group 2) admitted from January 2007 to April 2008 were selected to verify the expression of different genes, in which up-regulated genes including ESM1, MMP-10, SPP1 and HMGB1 were tested by quantitative real-time PCR and immunohistochemistry. RESULTS: Microarray analyses revealed 317 genes that exhibited > 2-fold or < 0.5 differences were identified (including 191 genes up-regulated and 126 down-regulated). Pathway analysis showed that these genes involved cell cycle, cell adhesion molecules, and extracellular matrix-receptor interaction were obviously predominant. Of them, 97 up-regulated genes and 44 down-regulated genes were related to angiogenesis. The mRNA expression of ESM1, MMP-10, SPP1 and HMGB1 in endometrial cancer group 2 were 0.898, 3.890, 1.433 and 1.881, respectively. Positive expression of SPP1, MMP-10, ESM1 and HMGB1 was observed in endometrial cancer group 2. However, the SPP1, ESM1 and HMGB1 was negative expressed in control group 2. CONCLUSION: It shows that there are the different angiogenesis related genes between endometrial cancer and normal endothelium, which will provide insights into the anti-angiogenesis therapy for endometrial cancers.

Intensity-modulated radiation therapy for advanced cervical cancer: a comparison of dosimetric and clinical outcomes with conventional radiotherapy.

OBJECTIVE: The aim of this study is to evaluate the dosimetry, efficacy and toxicity of reduced field intensity-modulated radiation therapy (RF-IMRT) for patients with advanced cervical cancer. METHODS: From August 2005 to August 2010, 60 patients with stage IIB-IIIB cervical cancer underwent reduced field IMRT (RF-IMRT group) and 62 patients treated with conventional radiotherapy (c-RT group) were enrolled. The RF-IMRT plans were as follows: whole pelvic IMRT plan was performed to deliver a dose of 30Gy firstly, then the irradiated volume was reduced to lymphatic drainage region as well as paracervix and parametrium for an additional 30Gy boost. Intracavitary brachytherapy and concurrent chemotherapy were performed during external irradiation. The tumor coverage and normal tissue avoidance were evaluated. Treatment response, toxicities and survival were assessed. RESULTS: The mean dose delivered to the planning target volume was significantly higher in RF-IMRT group than in c-RT group (61.5 vs. 50.8Gy, P=0.046). IMRT plans yielded better dose conformity to the target and better sparing of the rectal, bladder and small intestine. The RF-IMRT patients experienced significantly lower acute and chronic toxicities with comparable short-term effects than did those treated with conventional RT (CR: 87.7% vs. 88.3%, P=0.496; PR: 7.0% vs. 6.7%, P=0.440). No significant differences were found between treatment groups for 1year, 3year, and 5year overall survival (OS) levels, although the latter approached statistical significance in favor of IMRT, while a significantly higher progression-free survival (PFS; P=0.031) was seen for IMRT. CONCLUSIONS: RF-IMRT yields improved dose distributions, with lower toxicities, while providing comparable clinical outcomes. The increased PFS may be an advantage.

Sentinel lymph node biopsy as guidance for radical trachelectomy in young patients with early stage cervical cancer.

BACKGROUND: The purpose of this study was to assess the feasibility and accuracy of sentinel lymph nodes (SLNs) detection using 99mTc phytate in predicting pelvic lymph nodes status for radical abdominal trachelectomy (RAT) in patients with early stage cervical cancer. METHODS: Sixty-eight women with stage IA2-IB1 cervical cancer and scheduled to undergo fertility-sparing surgery enrolled in this study. 99mTc-labeled phytate was injected before surgery. Intraoperatively, SLNs were identified, excised, and submitted to fast frozen section. Systematic bilateral pelvic lymphadenectomy and/or para-aortic lymph node dissection was performed. Then RAT was performed in patients with negative SLNs. All nodes were sent for routine pathological examination and immunostained with anti-cytokeratin antibody to detect micrometastases. Outcomes of follow up and fertility were observed. RESULTS: SLNs were identified in 64 of 68 patients (94.1%). Of these, SLNs of 8 patients (11.8%) were positive on frozen sections and proved to be metastasis by final pathologic examination. The sensitivity, accuracy, and false negative rates were 100%, 100%, and 0%, respectively. All 60 patients with negative SLN underwent RAT successfully. Two relapses occurred and no one died of tumor progression during follow-up. Five of the 15 patients with procreative desire conceived 8 pregnancies (3 term delivery, 2 premature birth, 1 spontaneous abortion, and 2 were still in the duration of pregnancy) after surgery. CONCLUSIONS: The identification of SLN using 99mTc-labeled phytate is accurate and safe to assess pelvic nodes status in patients with early cervical cancer. SLNs biopsy guided RAT is feasible for patients who desire to have fertility preservation.

[Characteristics of foliar delta13C values of common shrub species in various microhabitats with different karst rocky desertification degrees].

By measuring the foliar delta13C values of 5 common shrub species (Rhamnus davurica, Pyracantha fortuneana, Rubus biflorus, Zanthoxylum planispinum, and Viburnum utile) growing in various microhabitats in Wangjiazhai catchment, a typical karst desertification area in Guizhou Province, this paper studied the spatial heterogeneity of plant water use at niche scale and the response of the heterogeneity to different karst rocky desertification degrees. The foliar delta13C values of the shrub species in the microhabitats followed the order of stony surface > stony gully > stony crevice > soil surface, and those of the majority of the species were more negative in the microhabitat soil surface than in the others. The foliar delta13C values decreased in the sequence of V. utile > R. biflorus > Z. planispinum > P. fortuneana > R. davurica, and the mean foliar delta13C value of the shrubs and that of typical species in various microhabitats all increased with increasing karst rocky desertification degree, differed significantly among different microhabitats. It was suggested that with the increasing degree of karst rocky desertification, the structure and functions of karst habitats were impaired, microhabitats differentiated gradually, and drought degree increased.

Intensity-modulated radiation therapy versus para-aortic field radiotherapy to treat para-aortic lymph node metastasis in cervical cancer: prospective study.

AIM: To compare dosimetry, efficacy, and toxicity of intensity-modulated radiation therapy (IMRT) with para-aortic field radiotherapy in patients with para-aortic lymph node (PALN) metastasis of cervical cancer. METHODS: This prospective study examined 60 patients with cervical cancer with PALN metastasis who underwent whole-pelvis radiotherapy followed by brachytherapy between November 1, 2004 and May 31, 2008. After 3 cycles of chemotherapy, patients were serially allocated into two groups and treated with IMRT or para-aortic field RT at doses of 58-68 Gy and 45-50 Gy, respectively. Treatment response was evaluated and toxicities were assessed. Patients in the IMRT group were treated with both para-aortic field RT and IMRT in order to compare the exposure dose of organs at risk. RESULTS: In the IMRT group, the mean dose delivered to the planning target volume was 67.5 Gy. At least 99% of the gross tumor volume received effective coverage and radical dose (median, 63.5 Gy; range, 54.5-66) during treatment. IMRT plans yielded better dose conformity to the target and better sparing of the spinal cord and small intestine than para-aortic field RT. The IMRT patients experienced less acute and chronic toxicities. The IMRT group also had higher 2- and 3-year survival rates than the para-aortic RT group (2-year, 58.8% vs 25.0%, P = 0.019; 3-year, 36.4% vs 15.6%, P = 0.016). However, no significant difference was found in 1-year survival (67.7% vs 51.3%, P =0.201). The median survival in the IMRT group was 25 months (range, 3 to 37 months). The actuarial overall survival, disease-free survival, and locoregional control rates at 2 years were 67%, 77%, and 88%, respectively, in the IMRT group. CONCLUSIONS: IMRT provides better clinical outcomes than para-aortic field radiotherapy in patients with PALN metastasis. However, cervical local and distal recurrence remain a problem. Long-term follow-up and studies involving more patients are needed to confirm our results.

[Gemcitabine based combination chemotherapy, a new salvage regimen for recurrent platinum resistant epithelial ovarian cancer].

OBJECTIVE: To evaluate the efficacy and toxicities of gemcitabine combined with ifosfamide and anthracycline chemotherapy for recurrent platinum resistant ovarian epithelial cancer. METHODS: Gemcitabine 800 mg/m(2) (day 1, 8), ifosfamide 1.5 g/m(2) (day 1 - 3), adriamycin 40 mg/m(2) or epirubicin 60 mg/m(2) (day 1) or mitoxantrone 10 mg/m(2) (day 1, 8) were used in recurrent platinum resistant/refractory ovarian cancer patients, the cycle was repeated at interval of 21 to 28 days. RESULTS: A total of 60 patients received 172 cycles combined chemotherapy. There were no one cases complete response, while partial response 22 (37%, 22/60), stable 23 (38%, 23/60) and progression 15 (25%, 15/60) were observed, with clinical benefit rate 75% (45/60). The median time of progression-free survival was 7 months, and the median overall survival time was 20 months. The main side effect was hematologic toxicity with leukopenia rate of 82% (49/60), among which III-IV accounted for 31% (15/49). Digestive reaction was all in I-II, accounted for 42% (25/60). CONCLUSION: The regimen of gemcitabine combined with ifosfamide and anthracycline is feasible, tolerable and effective in patients with recurrent platinum resistant/refractory epithelial ovarian cancer.

Inhibition of osteopontin suppresses in vitro and in vivo angiogenesis in endometrial cancer.

OBJECTIVE: Osteopontin (OPN) has been found to play an important role in tumor angiogenesis in recent years. Our previous studies have shown that OPN is overexpressed in tumor-associated human endometrial endothelial cells (HEECs) isolated from tissue samples of patients with endometrial cancer. In the present study, we aimed to further determine the role of OPN in endometrial cancer-associated angiogenesis. METHODS: We knock down OPN expression in HEECs and human endometrial cancer Ishikawa (ISK) cells using the small interference RNA method, and then evaluate the effects of OPN on endometrial cancer-associated angiogenesis by in vivo mouse studies and in vitro assays. RESULTS: Our results revealed that proliferative activity of HEECs and ISK cells in vitro was not affected by transfection with the siOPN-RNA (P>0.05). Inhibition of OPN expression in HEECs reduced the cell migration, with the percentage of repaired area of 36.32+/-2.88 vs. 8.54+/-1.13 (P=0.007). HEEC/siOPN and ISK/siOPN demonstrated 67.4% and 51.2% decreased invasiveness compared with controls, respectively (P<0.05). The number of branched points per well was obviously lower in HEEC/siOPN than that in HEEC/Control (32.46+/-17.10 vs. 53.15+/-15.44, P=0.021). Furthermore, ISK cells transfected with OPN siRNA formed smaller tumor in mice and led to a lower microvessel density, i.e., angiogenesis, in transplanted tumors of mice than scrambled siRNA controls (12.88+/-7.14 vs. 28.42+/-9.69 vessels per HPF, P=0.019). CONCLUSION: These data confirm the positive role of OPN in endometrial cancer-associated angiogenesis and might be of great benefit for finding rational approach in endometrial cancer therapy.

Silencing of heat shock protein 70 expression enhances radiotherapy efficacy and inhibits cell invasion in endometrial cancer cell line.

AIM: To investigate the role of heat shock proteins 70 (HSP70) in radiosensitivity and invasiveness of endometrial cancer in vitro. METHODS: HSP70 expression was silenced in relatively radioresistant, well-differentiated human endometrial cancer cell line ISK, using small interference RNA method, or by HSP70 overexpression after transfecting a HSP70-expressing vector. The effect of HSP70 on ISK cell line response to irradiation was evaluated. The surviving fraction was measured using colony-formation assay. Apoptosis was detected by flow cytometry and HSP70 expression was determined by quantitative real-time polymerase chain reaction, western-blot, and/or immunocytochemistry. Cell invasiveness was measured using transwell invasion assay. RESULTS: HSP70 silencing caused a significant increase in irradiation-induced cell killing in comparison with control cells, with an enhancement factor of 1.27, and in the percentage of apoptotic cells (14.22% vs 6.74%, P = 0.021). After 4 Gy irradiation, mean +/- standard deviation survival fraction in ISK cells was reduced to 0.32 +/- 0.04 in comparison with control values but in ISK/siRNA-HSP70 cells the survival fraction was higher and amounted to 0.51 +/- 0.08 (P = 0.026). Silencing HSP70 significantly inhibited cell invasion before and after irradiation (106 +/- 19 vs 219 +/- 18 and 119 +/- 16 vs 256 +/- 31, P = 0.007). On the contrary, ectopic overexpression of HSP70 attenuated irradiation-induced apoptosis (7.15% vs 4.08%, P = 0.043) and induced more ISK/HSP70 cells invaded through the filters than mock-infected cells before and after irradiation (274 +/- 21 vs 194 +/- 16 before irradiation, and 298 +/- 24 vs 227 +/- 19 after irradiation, respectively, P = 0.032). CONCLUSION: Disruption of HSP70-induced cytoprotection during irradiation enhances therapeutic effect of irradiation, which makes HSP70 a promising target in the research of endometrial cancer.

Differential expression profiling of gene response to ionizing radiation in two endometrial cancer cell lines with distinct radiosensitivities.

Although radiotherapy is routinely administered to high-risk endometrial carcinoma and offer a significant disease-free survival advantage, the therapeutic effect is sometimes limited by the occurrence of radioresistance. To determine the patterns of gene expression responsible for the radioresistance and to search for potential target genes for radiotherapy, we selected two cell lines with distinct radiosensitivities using colony-formation assay from four endometrial cancer cell lines. The cell cycle distribution showed higher fractions of G2/M phase cells in the radiosensitive cell line KLE after radiation compared with the radioresistant cell line ISK. Apoptosis assessment also showed significant elevation in the percentage of early apoptosis cells in KLE cells. Subsequently, gene expression changes after X-ray exposure were analyzed by using oligonucleotide microarrays. We identified, respectively, in ISK and KLE, 227 and 354 genes that exhibited > or =2-fold difference. However, only 53 genes showing differences more than double the median expression value between the two groups were defined as radiosensitivity (or radioresistance)-related genes. Among these, genes associated with DNA-repair, apoptosis, growth factor, signal transduction, cell cycle and cell adhesion were predominant. The validity of the expression level of 10 randomly selected genes was confirmed by real-time PCR and/or Western blotting. In conclusion, the differential gene expression changes that occur after radiation in the two cell lines will provide insight into molecular mechanisms of radioresistance in endometrial carcinoma, and also the means to find potential targets to achieve further gains in therapeutic benefit.

Gene alterations in tumor-associated endothelial cells from endometrial cancer.

The alterations in the gene expression profile of tumor-associated human endometrial endothelial cells (HEECs) may allow opportunities for developing new therapeutic approaches to inhibit angiogenesis in endometrial cancer. The aim of this study was to identify the different gene expression pattern between tumor-associated HEECs and normal HEECs. To elucidate the molecular mechanisms governing the abnormal vasculature in endometrial cancer, we examined global expression patterns of purified endothelial cells from three endometrial cancers and three age-matched normal endometria using oligonucleotide microarrays. We also performed in vitro culture and identified the endothelial origin, as well as observing the functional characteristics in angiogenesis, of HEECs from the two different sources. Microarray analyses revealed distinct gene expression patterns and consistent up-regulation of certain endometrial endothelial marker genes across patient samples. More than 300 genes that exhibited > or =2-fold differences were identified in tumor-associated HEECs. Pathway analysis showed that pathways of Cell cycle, Cell adhesion molecules (CAMs), focal adhesion, and extracellular matrix (ECM)-receptor interaction were obviously predominant. The results of the microarray analysis were confirmed by quantitative real-time PCR, immunohistochemistry, and/or Western blotting. Moreover, although the tumor-associated HEECs did not show faster proliferation than normal HEECs, they exhibited enhanced migration ability, potent invasiveness, and elevated tube formation in vitro. The present study shows that tumor and normal endothelium differ at the molecular level, and additional characterization of this gene expression database will provide insights into the angiogenesis of endometrial cancers and might be of great benefit for finding potential therapeutic targets.

Inhibition of lysophosphatidic acid receptor-2 expression by RNA interference decreases lysophosphatidic acid-induced urokinase plasminogen activator activation, cell invasion, and migration in ovarian cancer SKOV-3 cells.

AIM: To explore the role of lysophosphatidic acid receptor-2 (LPA2) in regulating lysophosphatidic acid (LPA)-induced urokinase plasminogen activator (uPA) activation, cell invasion, and migration in human ovarian cancer cell line SKOV-3. METHODS: SKOV-3 cells were stimulated with LPA. Cell supernatant uPA level and activity were measured using enzyme-linked immunosorbent assay. LPA2 mRNA expression was inhibited with LPA2-specific small interfering RNA (siRNA) and examined using semiquantitative reverse transcriptase-polymerase chain reaction. LPA-induced cell invasion and migration in transfected cells were evaluated by a Matrigel invasion chamber and a Transwell chemotaxis chamber, respectively. RESULTS: LPA stimulation significantly enhanced in vitro uPA activity in time- and dose-dependent manner. The levels of LPA-induced uPA protein decreased by 55% in LPA2 siRNA-transfected cells compared with negatively transfected cells at 24 hours after being treated with 80 micromol/L LPA (0.75+/-0.03 vs 0.34+/-0.04, P=0.004). In the LPA2 specific siRNA-transfected SKOV-3 cells, LPA treatment at 80 micromol/L induced considerably less invasion and migration compared with negative control siRNA-transfected SKOV-3 cells (invasion: 178+/-17.2 vs 36.2+/-3.3, P=0.009; migration: 220.4+/-25.5 vs 57+/-7.6, P=0.009). CONCLUSION: LPA2 has an essential role in LPA-induced uPA activation and tumor cell invasion in ovarian cancer SKOV-3 cells.

[Responses of delta13 C values of plant leaves to environmental gradients along environmental gradient factors in rocky desertified area of a typical karst Ggorge].

We analyzed the responses of delta13 C values of plant leaves to environmental factors (namely, soil water storage, air relative humidity, light intensity, depths of soil, soil organic content, average temperature and soil water content) and the correlations between them, by measuring delta13 C values of leaves for 11 plants species from 4 typical communities with different karst rocky desertification backgrounds in a typical karst catchments basin, Huajiang Gorge. It is revealed that, the delta13 C values and water use efficient of most species decrease with the increasing of water supply; but a few species exhibit an opposite trend and several others exhibit no change in delta13 C values or water use efficiency when these environmental factors varied. Moreover, the correlation analysis indicates that the soil water storage is the leading factor for Pistacia weinmannifolia, Mallotus repandus and Alchornea trewioides, while the depths of soil is essential factor for Nephrolepis cordifolia and Mallotus japonicus var. floccosus, and the light intensity is leading factor for N. cordifolia, Alangium chinense, Broussonetia papyrifera. However, the leading factor for some species like Rapanea kwangsiensis, Sapium rotundifolium and Cipadessa cinerascens are yet not clear, which mean their delta13 C values are affected by more comprehensive factors. Hence it could be concluded that high delta13 C values of leaves could indicate the adaptability of plants for low water regime, high light and low resource environments.