Guiling Li

HPV E6 down-regulation and apoptosis induction of human cervical cancer cells by a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott in vitro.

AIM OF THE STUDY To evaluate the cytotoxicity and apoptosis induction effects of a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott on CaSki, HeLa and HBL-100 cells. Particularly, the effect of PE on HPV E6 gene expression was tested, and the mechanism of its apoptosis induction effect was also studied. MATERIALS AND METHODS Cell viability was measured by the MTT assay. DAPI staining and flow cytometric analysis (FCM) were used to identify apoptotic cells in PE-treated CaSki, HeLa, and HBL-100 cells. Expression of the HPV E6 gene in CaSki and HeLa cells was detected by real-time RT-PCR and western blot analysis. Apoptosis-associated genes were examined by RT-PCR and western blot analysis in CaSki cells. RESULTS PE inhibited the growth of CaSki and HeLa cells in a time- and dose-dependent manner, but it had no obvious inhibiting effect on HBL-100 cells except at a relatively high dose (500 μg/mL). PE could induce apoptosis in CaSki and HeLa cells in a time-dependent manner but not in HBL-100 cells. HPV E6 mRNA and protein were decreased significantly by PE. Caspase-8, caspase-3, Bax, P53 and P21 mRNAs as well as proteins were increased while Bcl-2 mRNA and protein were decreased significantly by 24 h of PE treatment. CONCLUSIONS PE can function as a tumor suppressor by inducing apoptosis in human cervical cancer cells but it has little side effect on normal cells. It probably acts via mitochondria-dependent and death receptor-dependent apoptotic pathways. HPV E6 may be the key target of its action.

Ovarian Masses in Children and Adolescents - An Analysis of 521 Clinical Cases

OBJECTIVE To analyze the clinical characteristics of ovarian masses in children and adolescents. MATERIALS AND METHODS We performed a retrospective analysis of patients less than 20 years of age who were treated at the Obstetrics and Gynecology Hospital of Fudan University between March 2003 and January 2012. Medical records were reviewed for age at operation, including presentation of symptoms and signs; the levels of tumor markers; imaging examinations; pathologic findings; the size of masses; treatment; and outcome. Data management and descriptive analyses were performed using SPSS 16.0. RESULTS A total of 521 patients were included in this study. Among them, 92 had non-neoplastic lesions, 382 had benign neoplasms, and 47 had malignant tumors. The mean age of the patients was 16.3 ± 2.2 years. The primary presenting symptoms and signs were abdominal pain (39.5%), menstrual disorder (31.1%), abdominal swelling (5.4%), and an enlarged abdominal perimeter (3.3%). Malignant tumors tended to be larger than benign neoplasms (17.3 ± 8.6 cm vs 9.0 ± 5.7 cm; P = .000). There was no age difference between patients with benign neoplasms (16.3 ± 2.1 y) and those with malignant tumors (15.7 ± 2.5 y). The operations included salpingo-oophorectomy, ovarian cystectomy, and oophorectomy. Two patients with malignant tumors had bilateral salpingo-oophorectomy, and 2 patients who had tumor metastasis underwent a total abdominal hysterectomy and bilateral salpingo-oophorectomy. Forty-one cases of malignant tumors received postoperative chemotherapy. CONCLUSIONS Germ cell tumors are the most common malignancy, and mature teratomas are the most common benign neoplasms in children and adolescents. Abdominal pain and menstrual disorder are the main reasons for doctors visit. Although examination by ultrasound is the preferred auxiliary in the diagnosis of ovarian pathology, it could not distinguish between benign and malignant tumors. However, tumor size and tumor markers are helpful to identify the properties of masses. Surgery is usually better for treatment, and it is preferable to attempt conservative, fertility-sparing surgery in adolescents. Postoperative chemotherapy is necessary for malignant tumors.

Minimal deviation adenocarcinoma of the uterine cervix

Precise preoperative diagnosis of minimal deviation adenocarcinoma (MDA) of the uterine cervix is often difficult because the histological features of MDA closely resemble those of normal cervical glands.

Synergistic effects of a novel lipid-soluble extract from Pinellia pedatisecta Schott and cisplatin on human cervical carcinoma cell lines through the regulation of DNA damage response signaling pathway

Herbal medicines have been recognized as an attractive approach for cancer therapy with minimal side effects. The present study investigated the type of interaction between a novel lipid-soluble extract from Pinellia pedatisecta Schott (PE) and cisplatin (CDDP) on human cervical cancer SiHa and CaSki cell lines in vitro. The mechanism of this combination was studied using cell proliferation, invasion and apoptosis assays, and by analyzing cell cycle distribution and protein expression, with a focus on DNA damage response (DDR) activation. Equipotent combinations of PE and CDDP were determined by isobologram analysis, in order to evaluate potential synergy. The combination index for SiHa cells was 0.43, and the index for CaSki cells was 0.68, indicating synergy. Treatment with PE and CDDP combined resulted in a significantly greater inhibition of invasion in the two cells, compared with either drug alone (SiHa, P<0.01; CaSki, P<0.001). This co-treatment induced significantly more apoptosis in the two cell lines, and arrested cells at the G0/G1 phase and G2/M phase in SiHa and CaSki, respectively, with a significant decrease (P<0.01) in S phase cells in the two cell lines. Combined PE and CDDP targeting synergistically enhanced the expression of markers of DDR (phosphorylation of ataxia-telangiectasia mutated, checkpoint kinase (Chk)-1, Chk-2, and γ-H2A histone family member X) in cells. These results suggest that PE and cisplatin act synergistically in cervical cancer cells with high DDR activation. The approach presented in the present study may have important implications for the pharmacological mechanism of Pinellia pedatisecta Schott and cervical cancer therapeutic strategies.

Synergistic cytotoxic effects of a combined treatment of a Pinellia pedatisecta lipid-soluble extract and cisplatin on human cervical carcinoma in vivo

Herbal medicines are known to have numerous benefits, including lower toxicity and fewer side effects than traditional chemotherapeutic drugs. In traditional Chinese medicine, the rhizome of Pinellia pedatisecta (PE) Schott has long been used to treat cancer, undiagnosed swelling and erythema toxicum. However, its medical benefits lack support from scientific evidence. A novel lipid-soluble extract from PE has been previously verified to enhance the cytotoxicity of cis-dichlorodiammineplatinum-II (CDDP) against human cervical cancer cells in vitro. The present study evaluated the synergistic cytotoxic effects of PE and CDDP against human cervical cancer. Combination therapy of PE with CDDP exhibited synergistic cytotoxicity towards CaSki cell growth in mouse xenograft tumors. PE exhibited a cytotoxic effect on tumor size and weight, although the inhibitory ratio of tumor weight was only 26.3% in the PE-treated group. However, when mice were co-treated with PE and CDDP, the inhibitory ratio was higher than that of mice treated with CDDP alone (50.8 vs. 68.4%, respectively). The potential synergistic mechanism was likely via inhibiting the signaling E6/p53 pathway, restoring p53 function and inducing downstream tumor suppressor chain effects on apoptosis. Western blot analysis and immunohistochemistry indicated thatE6protein expression was significantly decreased upon treatment with combined PE and CDDP. The expression of p53 was increased in the combined PE and CDDP treatment group. Upregulation of p53-dependent apoptosis-associated proteins, including Bcl-2-associated X protein and cleaved caspases-9 and −3, was observed in the combined PE and CDDP treatment group. Our results present a molecular basis for the future application of the combination of PE and CDDP in the treatment of cervical cancer as a novel and pharmacologically safe chemotherapeutic strategy.

Delineation of retroperitoneal metastatic lymph nodes in ovarian cancer with near‑infrared fluorescence imaging

Lymph node metastasis occurs in early-stage and late-stage ovarian cancers. Systematic lymphadenectomy is frequently conducted in an attempt to prevent disease progression. However, this method is associated with multiple complications. Therefore, it is necessary to develop a less invasive and more sensitive method for detecting lymphatic metastasis in ovarian cancer. The aim of the present study was to develop an appropriate fluorescent label for the analysis of lymphatic metastasis in vivo. To this end, epithelial ovarian cancer cells with high potential for lymph node metastasis were labeled using mCherry fluorescence. The cells were then imaged in vitro to determine the expression of mCherry, and in a mouse xenograft model in vivo. The data demonstrated the successful identification of metastatic retroperitoneal lymph nodes by co-localization with lymph nodes labeled by near-infrared fluorescence nanoparticles in vivo. These data provided important insights into the further development of methods for intra-operative identification of lymphatic metastasis and the mechanisms underlying lymphatic metastasis.

Identification of lymphatic metastasis-associated genes in a metastatic ovarian cancer cell line.

Ovarian cancer (OC) is the gynecological malignancy with the highest rate of mortality, and lymphatic metastasis is a critical factor in disease recurrence and prognosis. In the current study, the SKOV-3/LN403 cell line, which has high potential for lymph node metastasis was established as a result of four rounds of selection from retroperitoneal lymph nodes following intraperitoneal injections of SKOV-3 ovarian adenocarcinoma cells. In comparison to the parental SKOV-3 cell line, SKOV-3/LN403 has a higher rate of proliferation, is more invasive and exhibits greater resistance to paclitaxel. Subsequently, a novel animal model of OC lymphatic metastasis was developed with SKOV-3/LN403 cells and a high incidence of positive metastatic lymph nodes, peritoneal dissemination and bloody ascites were observed, which mimicked the clinical outcome of patients with OC. Analysis of the gene expression profiles of SKOV-3 and SKOV-3/LN403 cells identified several genes and pathways that may be involved in lymphatic metastasis of OC. The induction of focal adhesion kinase expression provides a potential therapeutic target for OC lymphatic metastasis.

Immune modulation of a lipid-soluble extract of Pinellia pedatisecta Schott in the tumor microenvironment of an HPV+ tumor-burdened mouse model

ETHNOPHARMACOLOGICAL RELEVANCE Pinellia pedatisecta Schott extract (PE), a traditional Chinese medicine, has been used to reduce swelling, dry dampness and suppress cervical tumors. AIMS To evaluate the roles of PE in the regulation of anti-tumor effects and the cellular immune response in the tumor microenvironment. METHODS The immune microenvironment of HPV+TC-1 tumors was examined by immunohistochemistry, real-time PCR and flow cytometry. RESULTS Our study demonstrated that PE in vitro could significantly increase the percentage of apoptosis and necrosis in HPV+TC-1 cells and block the cell cycle phase. In vivo treatment with PE eradicated established subcutaneous HPV+TC-1 tumors in wild-type C57BL/6 mice by infiltrating CD8+ T cells and CD4+ T cells and by directly suppressing tumor growth and resistance to avascular necrosis. The key factors in the canonical Wnt signaling pathway in the experimental group (PE+mDC+naive CD4+T cells) were challenged, and the levels of beta-catenin, C-myc, cyclin D1 and PPAR1 were significantly enhanced at the 5th day. In particular, the subset proportion of Th1 cells (characterized by IFNγ production and the transcription factor Tbet) increased significantly, and both Th2 cells (characterized by IL-4 production and the transcription factor GATA3) and Th17 cells (characterized by IL-17 production and the transcription factor RoRγt) decreased profoundly. CONCLUSIONS These findings linked the anti-tumor properties of PE with the immune microenvironment to present a reliable basis for the future practical application of PE in cervical cancer as a novel and pharmacologically safe immunotherapy strategy.

Congenital vaginal atresia: A report of 39 cases in a regional Obstetrics and Gynecology Hospital

To investigate the clinical course and management of congenital vaginal atresia. This retro-spective analysis included patients with congenital vaginal atresia treated from March 2004 to August 2014 at the Obstetrics and Gynecology Hospital of Fudan University. Thirty-nine patients were included in this study. Their average age was 16.87±2.2 years when they came to our hospital. Totally, 51% of the patients had isolated congenital vaginal atresia with a normal cervix, whereas the others had either cervical atresia or imperforate hymen. The primary presenting signs and symptoms included primary amenorrhea (71.8%), periodic abdominalgia (41.0%), abdominal pain (36.0%), dyspareunia (10.3%), menstrual disorders (5.1%), and pelvic mass (5.1%). Ultrasound and magnetic resonance imaging (MRI) were effective inspection methods for the screening of urogenital tract-associated anomalies. Vagi-noplasty mainly included simple vagina reconstruction with insertion of a mold (n=22) and split-thickness skin grafting (n=4). In 64% of surgical patients, normal menstrual bleeding was achieved. Four of the patients subsequently became pregnant and delivered at term. Primary amenorrhea, periodic abdominalgia and abdominal pain are the main reasons for the post pubertal patients to visit doctors. Surgical methods can successfully provide these patients an opportunity for subsequent conservative management, can result in normal menstrual bleeding, resolve cyclic pelvic pain, and provide some po-tential for fertility.SummaryTo investigate the clinical course and management of congenital vaginal atresia. This retro-spective analysis included patients with congenital vaginal atresia treated from March 2004 to August 2014 at the Obstetrics and Gynecology Hospital of Fudan University. Thirty-nine patients were included in this study. Their average age was 16.87±2.2 years when they came to our hospital. Totally, 51% of the patients had isolated congenital vaginal atresia with a normal cervix, whereas the others had either cervical atresia or imperforate hymen. The primary presenting signs and symptoms included primary amenorrhea (71.8%), periodic abdominalgia (41.0%), abdominal pain (36.0%), dyspareunia (10.3%), menstrual disorders (5.1%), and pelvic mass (5.1%). Ultrasound and magnetic resonance imaging (MRI) were effective inspection methods for the screening of urogenital tract-associated anomalies. Vagi-noplasty mainly included simple vagina reconstruction with insertion of a mold (n=22) and split-thickness skin grafting (n=4). In 64% of surgical patients, normal menstrual bleeding was achieved. Four of the patients subsequently became pregnant and delivered at term. Primary amenorrhea, periodic abdominalgia and abdominal pain are the main reasons for the post pubertal patients to visit doctors. Surgical methods can successfully provide these patients an opportunity for subsequent conservative management, can result in normal menstrual bleeding, resolve cyclic pelvic pain, and provide some po-tential for fertility.