Assisted reproductive technology and physical activity among Chinese pregnant women at high risk for gestational diabetes mellitus in early pregnancy: A cross-sectional study.

Currently, the number of pregnant women at high risk for gestational diabetes mellitus (GDM) and using assisted reproductive technology (ART) is increasing. The present study aims to explore the relationship between ART and physical activity in Chinese pregnant women at high risk for GDM in early pregnancy. A cross-sectional study was conducted in a regional teaching hospital in Guangzhou, China, between July 2022 and March 2023. Three hundred fifty-five pregnant women at high risk for GDM in early pregnancy completed the Chinese version of the Pregnant Physical Activity Questionnaire (PPAQ), the Pregnancy Physical Activity Knowledge Scale, the Pregnancy Physical Activity Self-Efficacy Scale, the Pregnancy Physical Activity Social Support Scale, and a sociodemographic and obstetric characteristics data sheet. Compared to women who conceived naturally, women who used ART were more likely to be 35 years or older, unemployed, primigravidae, and to have intentionally planned their pregnancies. Women who used ART had significantly lower levels of physical activity and self-efficacy compared to their counterparts who conceived naturally. Over half (55.6%) of women who used ART reported being physically inactive, and those with lower self-efficacy, as well as the unemployed, were significantly more likely to be inactive. Physical inactivity is a critical clinical issue among women who use ART, especially in the context of GDM risk. Future research should develop and test physical activity programs, including enhancing physical activity self-efficacy for women who use ART. Patient or public contribution: In this study, survey questionnaires were completed by participants among Chinese pregnant women at high risk for GDM in early pregnancy.

Secondhand smoking exposure and quality of life among pregnant and postnatal women: a network approach.

OBJECTIVE: This study examined the prevalence of exposure to secondhand smoke, its correlates and its association with quality of life (QOL) among pregnant and postnatal Chinese women. DESIGN: This was a multicentre, cross-sectional study. SETTING: Participants were consecutively recruited from eight tertiary hospitals located in eight municipalities or provinces in China. PARTICIPANTS: A total of 1140 women were invited to join this study and 992 (87.02%) completed all measures. PRIMARY AND SECONDARY OUTCOME: Measures women's secondhand smoking behaviour (frequency and location of exposure to secondhand smoking), and their QOL measured by the WHO Quality of Life Questionnaire. RESULTS: A total of 211 women (21.3%, 95% CI 18.7% to 23.8%) had been exposed to secondhand smoking. Exposure to secondhand smoking was most common in public areas (56.4%), and residential homes (20.5%), while workplaces had the lowest rate of exposure (13.7%). Women with physical comorbidities were more likely to report secondhand smoking exposure, while older women, women living in urban areas, those with college or higher education level, and women in their second trimester were less likely to report exposure to secondhand smoking. Network analysis revealed that there were six significant links between secondhand smoke and QOL items. The strongest negative edge was the connection between secondhand smoke and QOL9 ('physical environment health', edge weight=-0.060), while the strongest positive edge was the connection between secondhand smoke and QOL3 ('pain and discomfort', edge weight=0.037). CONCLUSION: The prevalence of exposure to secondhand smoking is becoming lower among pregnant and postnatal women in China compared with findings reported in previous studies. Legal legislation should be maintained and promptly enforced to establish smoke-free environments in both public and private urban/rural areas for protection of pregnant and postnatal women, especially those who are physically vulnerable and less educated.

Extraction of antibacterial peptides against Helicobacter pylori from bovine milk casein.

Introduction: More than half of the world's population is infected with Helicobacter pylori, which may cause gastritis, peptic ulcer and even gastric cancer. The World Health Organization (WHO) has announced that H. pylori infection is a class I carcinogen and hence eradication of it is highly important. Bovine milk contains caseins, which can be digested by various enzymes in the human stomach to produce antibacterial peptides. Material and methods: This study used in vitro methods to extract anti-H. pylori peptides from caseins by the gastric protease pepsin under environments with similar pH values to those found in the human stomach. The molecular weights and sequences of the peptides were identified by MALDI-TOF mass spectrometry and MS/MS Ion Search, respectively. Antibacterial activity tests were performed to calculate the minimum inhibitory concentration (MIC90) of the extracts. Results: The findings of this study revealed that the major products of bovine milk casein digestion by pepsin are casecidin 17 and ß-casein 207-224. The extracts produced promising anti-H. pylori effects with the lowest MIC90 found at pH values of 1.5 and 2.0. Conclusions: This study identified the anti-H. pylori effects of casecidin 17 and ß-casein 207-224, which may help in developing therapeutic agents to modulate the effect of antibiotics on H. pylori infections.

Erratum: MicroRNA-125b Down-Regulation Mediates Endometrial Cancer Invasion by Targeting ERBB2.

The Figure 2 and Figure 4C were incorrectly published in the article titled MicroRNA-125b down-regulation mediates endometrial cancer invasion by targeting ERBB2. Chao Shang, Yan-ming Lu, Li-rong Meng, Med Sci Monit 2012; 18(4): BR149-155. 10.12659/MSM.882617. The correct Figures are as follows.

In Silico and In Vitro Anti-Helicobacter Pylori Effects of Combinations of Phytochemicals and Antibiotics.

Helicobacter pylori infection is a WHO class 1 carcinogenic factor of gastric adenocarcinoma. In the past decades, many studies have demonstrated the increasing trend of antibiotic resistance and pointed out the necessity of new effective treatment. This study was aimed at identifying phytochemicals that can inhibit H. pylori and possibly serve as adjuvant treatments. Here, in silico molecular docking and drug-like properties analyses were performed to identify potential inhibitors of urease, shikimate kinase and aspartate-semialdehyde dehydrogenase. These three enzymes are targets of the treatment of H. pylori. Susceptibility and synergistic testing were performed on the selected phytochemicals and the positive control antibiotic, amoxicillin. The in-silico study revealed that oroxindin, rosmarinic acid and verbascoside are inhibitors of urease, shikimate kinase and aspartate-semialdehyde dehydrogenase, respectively, in which, oroxindin has the highest potency against H. pylori, indicated by a minimum inhibitory concentration (MIC) value of 50 µg/mL. A combination of oroxindin and amoxicillin demonstrated additive effects against H. pylori, as indicated by a fractional inhibitory concentration (FIC) value of 0.75. This study identified phytochemicals that deserve further investigation for the development of adjuvant therapeutic agents to current antibiotics against H. pylori.

Sleep disturbance and its relationship with quality of life in older Chinese adults living in nursing homes.

PURPOSE: To examine sleep disturbances in older adults in Macau and Guangzhou, China and their associated factors. DESIGN AND METHODS: Four-hundred and thirty seven subjects in Guangzhou and 244 subjects in Macau were interviewed. FINDINGS: In total, 681 older adults participated in the study, and 27.8% reported sleep disturbance, with 43.9% in Macau and 18.8% in Guangzhou. Physical quality of life was negatively associated with sleep disturbances. Severe depressive symptoms were positively related but living in Guangzhou was negatively related to sleep disturbances. Sleep disturbances are more common in Macau compared to Guangzhou. PRACTICE IMPLICATIONS: Appropriate screening and treatment strategies are needed to address sleep disturbance in this population.

Elder abuse and its impact on quality of life in nursing homes in China.

There are limited available data on elder abuse and its impact on quality of life (QOL) in China. This study investigated the prevalence of elder abuse in nursing homes and its associated demographic, clinical factors and QOL in Macau and Guangzhou, China. A total of 681 subjects (244 in Macau and 437 in Guangzhou) were consecutively recruited. The prevalence of elder abuse was 11.48% and 8.24% in Macau and Guangzhou, respectively. Multivariate analyses revealed that having a religion and depressive symptoms were independently and positively associated with elder abuse. No significant association between elder abuse and any QOL domain was found. Elder abuse is common in nursing homes in both Macau and Guangzhou. Appropriate strategies and educational programs should be developed for health professionals to reduce the risk of elder abuse.

Clinical characteristics and quality of life of older adults with cognitive impairment in Macao.

BACKGROUND: Little is known about the characteristics of older adults with cognitive impairment in Macao. This study aimed to determine the prevalence of cognitive impairment and the quality of life (QOL) of older adults living in the community and nursing homes. METHODS: A consecutive sample of 413 subjects (199 from the community; 214 from nursing homes) was recruited and interviewed using standardized instruments. Cognition was measured with the Repeatable Battery for the Assessment of Neuropsychological Status and QOL with the brief version of the World Health Organization Quality of Life instrument. RESULTS: Altogether 87 subjects (21.0%) had cognitive impairment. On multivariate analyses, advanced age (P < 0.001, OR = 1.06, 95%CI: 1.03-1.1) and depressive symptoms (P = 0.03, OR = 1.07, 95%CI: 0.005-1.1) were positively associated with cognitive impairment. Married marital status (P = 0.01, OR = 0.3, 95%CI: 0.1-0.7) and higher education level (P < 0.001, OR = 0.1, 95%CI: 0.06-0.3) were negatively associated with cognitive impairment. After the confounders were controlled for, cognitive impairment was significantly associated with the lower psychological (F (11,412) = 6.3, P = 0.01) and social relationship domains of QOL (F (11,412) = 4.0, P = 0.04). CONCLUSION: Cognitive impairment was found to be common in community-dwelling and nursing home resident older adults in Macao. Given cognitive impairment's negative impact on QOL, appropriate strategies should be implemented to improve access to treatment in this population.

Prevalence of Sleep Disturbances and Their Associations With Demographic and Clinical Characteristics and Quality of Life in Older Adults in Macao.

PURPOSE: To examine the prevalence of sleep disturbances (difficulty initiating sleep [DIS], difficulty maintaining sleep [DMS], and early morning awakening [EMA]), their socio-demographic and clinical correlates, and quality of life (QOL) in older adults in Macao. DESIGN AND METHODS: Four hundred fifty-one subjects were interviewed using standardized instruments. FINDINGS: The prevalence of at least one type of sleep disturbance was 38.1%; the figures of DIS, DMS, and EMA were 18.6, 31.3, and 23.9%, respectively. Female sex and depressive symptoms were independently associated with more frequent sleep disturbances. Sleep disturbances were independently associated with lower physical QOL. PRACTICE IMPLICATIONS: Sleep disturbances are common in older adults in Macao. Appropriate strategies should be implemented to prevent and treat sleep disturbances and concerted attempts should be made to improve access to treatment.

Memory Impairment Following Electroconvulsive Therapy in Chinese Patients with Schizophrenia: Meta-Analysis of Randomized Controlled Trials.

PURPOSE: To evaluate memory impairment associated with electroconvulsive therapy (ECT)-antipsychotic (AP) combination in comparison to AP monotherapy in schizophrenia. DESIGN AND METHODS: A systematic literature search of randomized controlled trial (RCTs) was performed. FINDINGS: Eleven RCTs that compared ECT-AP combination (n = 508) with AP monotherapy (n = 510) were analyzed. ECT-AP combination was associated with greater impairment than AP monotherapy in (1) endpoint memory quotient (MQ) of the Wechsler Memory Scale (WMS)-Revised at the end of the ECT course; and (2) picture recall, counting, recognition, and associative learning of the WMS. However, no group difference was found in MQ at 1 and 2 weeks post-ECT. PRACTICE IMPLICATIONS: The ECT-AP combination was associated with greater transient memory impairment compared to AP monotherapy.

Blocking NOTCH pathway can enhance the effect of EGFR inhibitor through targeting CD133+ endometrial cancer cells.

Although the molecular therapeutics targeting key biomarkers such as epithelial growth factor receptor (EGFR), PI3K/AKT/mTOR, and vascular endothelial growth factor (VEGF) shows some success in clinical trials, some internally existing challenges in endothelial cancer biology hinder the drug effects. One of the major challenges stems from cancer stem cell-derived drug resistance. CD133 positive cells are well believed as cancer stem cells (CSC) in endometrial cancers and NOTCH pathway plays a critical role in retaining CD133+ cells by promoting CSC self-renewal and chemoresistance. Here, we initiated a therapeutic strategy to improve effects of EGFR inhibition by targeting NOTCH pathway of CD133+ cells in endometrial cancers. We first detected and purified the CD133+ cell fraction in endometrial cancer cell line Ishikawa (IK), and validated activation of NOTCH pathway in the CD133+ cells that have higher proliferation rate and lower apoptosis rate, comparing to CD133- cells. Results of nude mouse xenograft experiments further demonstrated CD133+ cells retain higher tumorigenesis capacity than CD133- cells, indicating their tumor-initiating property. Last, we applied both NOTCH inhibitor DAPT and EGFR inhibitor AG1478 treatment on endometrial cancer lines IK and HEC-1A and the results suggested improvement effects of the combination therapy compared to the treatments of DAPT or AG1478 alone. These findings indicated targeting NOTCH pathway in CD133+ cells, combining with EGFR inhibition, which provides a novel therapeutic strategy for endometrial cancer diseases.

Quality of life and clinical correlates in older adults living in the community and in nursing homes in Macao.

AIM: There have been no previous studies of quality of life (QOL) in older adults in Macao. This study aimed to examine QOL in relation to the sociodemographic and clinical characteristics of adults aged ≥50 years in Macao. METHODS: A sample of 451 subjects (203 living in the community, 248 living in nursing homes) was interviewed using standardized instruments. Basic sociodemographic and clinical data including QOL were collected. RESULT: There were no significant differences between the community and nursing home groups in any of the QOL domains. Multiple linear regression analyses revealed that poor physical QOL was significantly predicted by severe depressive symptoms, insomnia, major medical conditions, unmarried status, and lower education ( F 11,438 = 26.2, P < 0.001), which accounted for 38.2% of the variance. Poor psychological QOL was significantly predicted by severe depressive symptoms and lower educational level ( F 11,438 = 24.3, P < 0.001), which accounted for 36.4% of the variance. Poor social QOL was significantly predicted by severe depressive symptoms, male gender, and unmarried status ( F 11,438 = 5.6, P < 0.001), which accounted for 12.5% of the variance. Poor environment QOL was significantly predicted by lower educational level, severe depressive symptoms, and younger age ( F 11,438 = 6.6, P < 0.001), which accounted for 12.1% of the variance. CONCLUSION: Older Macanese adults had poorer scores on physical and social QOL domains than the general Hong Kong Chinese population. Their QOL was more strongly related to severe depressive symptoms, major medical conditions, and insomnia.

Synergistic antibacterial effects of herbal extracts and antibiotics on methicillin-resistant Staphylococcus aureus: A computational and experimental study.

Antibiotic resistance has become a serious global concern, and the discovery of antimicrobial herbal constituents may provide valuable solutions to overcome the problem. In this study, the effects of therapies combining antibiotics and four medicinal herbs on methicillin-resistant Staphylococcus aureus (MRSA) were investigated. Specifically, the synergistic effects of Magnolia officinalis, Verbena officinalis, Momordica charantia, and Daphne genkwa in combination with oxacillin or gentamicin against methicillin-resistant (ATCC43300) and methicillin-susceptible (ATCC25923) S. aureus were examined. In vitro susceptibility and synergistic testing were performed to measure the minimum inhibitory concentration and fractional inhibitory concentration (FIC) index of the antibiotics and medicinal herbs against MRSA and methicillin-susceptible S. aureus. To identify the active constituents in producing these synergistic effects, in silico molecular docking was used to investigate the binding affinities of 139 constituents of the four herbs to the two common MRSA inhibitory targets, penicillin binding proteins 2a (PBP2a) and 4 (PBP4). The physicochemical and absorption, distribution, metabolism, and excretion properties and drug safety profiles of these compounds were also analyzed. D. genkwa extract potentiated the antibacterial effects of oxacillin against MRSA, as indicated by an FIC index value of 0.375. M. officinalis and V. officinalis produced partial synergistic effects when combined with oxacillin, whereas M. charantia was found to have no beneficial effects in inhibiting MRSA. Overall, tiliroside, pinoresinol, magnatriol B, and momorcharaside B were predicted to be PBP2a or PBP4 inhibitors with good drug-like properties. This study identifies compounds that deserve further investigation with the aim of developing therapeutic agents to modulate the effect of antibiotics on MRSA. Impact statement Antibiotic resistant is a well-known threat to global health and methicillin-resistant Staphylococcus aureus is one of the most significant ones. These resistant bacteria kill thousands of people every year and therefore a new effective antimicrobial treatment is necessary. This study identified the herbs and their associated bioactive ingredients that can potential the effects of current antibiotics. These herbs have long history of human usage in China and have well-defined monograph in the Chinese Pharmacopeia. These indicate their relatively high clinical safety and may have a quicker drug development process than that of a new novel antibiotic. Based on the results of this study, the authors will perform further in vitro and animal studies, aiming to accumulate significant data for the application of clinical trial.

Effect of mTOR inhibitors in nude mice with endometrial carcinoma and variable PTEN expression status.

BACKGROUND: The aim of this study was to investigate the sensitivity to rapamycin of endometrial cancer cells with different phosphatase and tensin homologue (PTEN) expression to understand the mechanism of resistance to mammalian target of rapamycin (mTOR) inhibitors in the treatment of endometrial cancer. MATERIAL AND METHODS: Twenty specific pathogen-free female BALB/c mice received transplants of either HEC-1A (PTEN-positive) or Ishikawa (PTEN-negative) cells. Mice in the treatment group were injected intraperitoneally once a week for 4 consecutive weeks. The control group was injected weekly with phosphate buffer saline (PBS) for 4 consecutive weeks. Tumor volume, tumor mass, growth curves, and inhibition rate were measured, after which the mice were killed. RESULTS: Both tumor growth rate and size were slower in the treatment group than in the control group for all mice that received transplants of either HEC-1A or Ishikawa cells. The tumor inhibition rates in the treatment group were 48.1% and 67.1% in mice transplanted with HEC-1A and Ishikawa cells, respectively. CONCLUSIONS: The inhibitory effects of rapamycin were enhanced in PTEN-negative Ishikawa tumor cells compared with PTEN-positive HEC-1A cells, which could explain the reduced effect of rapalogues in some endometrial cancer patients and help to understand the mechanism of resistance to this drug.

MicroRNA-125b down-regulation mediates endometrial cancer invasion by targeting ERBB2.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding nucleotides that regulate mRNA stability and protein expression by imperfect base pairing with the 3'-untranslated region (3'UTR) of target mRNAs. Many miRNAs have been documented to be aberrantly expressed in human cancers, but the role of miRNAs in endometrioid endometrial cancer (EEC) remains poorly understood. The objective of this study was to investigate the effect of miR-125b on EEC development and to explore its molecular mechanism in EEC carcinogenesis. MATERIAL/METHODS: Real-time quantitative PCR was applied to evaluate the expression level of miRNA-125b in EEC and normal endometrium (NE) samples. The invasion ability of miR-125b in EEC HEC1B cells was analyzed by Transwell assay after pre-miR-125b or anti-miR-125b transfection. For the invasion mechanism analysis of miR-125b on HEC1B cells, miRBase, TargetScan, miRanda and PicTar were used to predict the possible target gene of miR-125b. Luciferase activities assay, cotransfection and Western blot were used to reveal that the predicted target genes of miR-125b were direct and specific. RNA interference technology was used to confirm that the invasion inhibition of miR-125b was directly induced by ERBB2. RESULTS: Our study showed that miR-125b was down-regulated in human EEC specimens compared to that in NC specimens. Over-expression of miR-125b in HEC1B cells inhibited EEC invasion and this inhibitory effect on HEC1B cells could be restored by miR-125b knock down. Mechanism analysis revealed that ERBB2 was a direct and specific target of miR-125b. The inhibitory effect on EEC cell invasion was mediated by miR-125b inhibition of the translation of a proto-oncogene, ERBB2. CONCLUSIONS: Aberrantly expressed miR-125b contributes to HEC1B cells invasion partly through directly down-regulating ERBB2 protein expression in EEC. This miRNA signature offers a novel potential therapeutic strategy for EEC.

KCC1 gene advances cell invasion ability by regulating ERK signaling pathway in endometrial cancer HEC-1B cell line.

INTRODUCTION: Human potassium chloride cotransporter-1 (KCC1) gene is expressed in endometrial cancer and related to metastasis of endometrial cancer. However, whether KCC1 contributes to invasion and metastasis of endometrial cancer has not been thoroughly investigated. The purpose of this study is to research the alternation effect of insulin-like growth factor I (IGF-I) on the expression of KCC1 in endometrial cancer HEC-1B cells and to explore the mechanism of how KCC1 regulates the invasion ability of HEC-1B cells through the extracellular signal-regulated kinase (ERK) signaling pathway. METHODS: First, the inhibitive effect of RNA interference to KCC1 was detected by semiquantitative reverse transcriptase-polymerase chain reaction. Western blot was used to measure expression changes of KCC1 after exposure to IGF-I in the HEC-1B cells. The change in quantity of phosphorylated ERK1/2 (p-ERK1/2) and cell invasion ability also were measured. After RNA interference and treatment with U0126, the quantity of p-ERK1/2 and the cell invasion ability were measured again. RESULTS: After the application of IGF-I on the HEC-1B cells, the expression of KCC1 and p-ERK1/2 increased dramatically, and the cell invasion ability advanced. RNA interference could inhibit the expression of KCC1, and the quantity of p-ERK1/2 and the cell invasion ability decreased even under the effect of IGF-I. Furthermore, after treatment with U0126, the cell invasion ability no longer advanced even under the effect of IGF-I either. CONCLUSIONS: Insulin-like growth factors I can induce the upregulation of KCC1 gene, and KCC1 gene participates in the invasion ability of HEC-1B cells through the ERK signaling pathway.

[Influence of human epidermal growth factor receptor-2 siRNA on chemosensitivity to cisplatin of human ovarian carcinoma cells: an in vitro experiment].

OBJECTIVE: To investigate the effects of RNA interference (RNAi) targeting human epidermal growth factor receptor 2 (HER-2) gene on the change of chemosensitivity to cisplatin of ovarian carcinoma cells. Methods Three kinds of HER-2 gene targeting siRNA, HER-2 siRNA I-III, were synthesized and the best one (HER-2 siRNA III) was screened. Human ovarian carcinoma cells of the line SKOV3 were cultured randomly divided into 3 groups: HER-2 siRNA III group, transfected with HER-2 siRNA III, non-specific siRNA group, transfected with non-specific siRNA III, and control group, without transfection. Cisplatin of the concentrations of 0, 0.05, 0.2. 0.4, 0.8, 1.0, 2.0, 10, and 20 microg/ml was added into the culture fluid for 24 h. MTT method was used to detect the proliferation rate of the SKOV3 cells. Other SKOV3 cells were divided into 3 groups: siRNA group, transfected with HER-2 siRNA III, cisplatin group, exposed to cisplatin, and HER-2 siRNA III and exposed to cisplatin. Annexin V method and flow cytometry were used to detect the apoptosis of the SKOV3 cells. The HER-2 gene expression was assessed by Western blotting. The chemosensitivity of transfected cells to cisplatin was measured by MTT. Western blotting was used to detect the protein expression of apoptosis related proteins: Bcl-2, surviving, XIAP, and Smac. RESULTS: After exposed to cisplatin, the cell survival rate decreased as the dose of cisplatin increased. The proliferation rate of the SKOV3 cells transfected with HER-2 siRNA III and exposed to 1 microg/ml cisplatin was (58 +/- 5)%, significantly lower than those of the nonspecific siRNA III transfection group [(65 +/- 6)%] and the control group [(68 +/- 3)%, both P < 0.01]. No significant difference in the cell survival rate was found between the control and nonspecific groups (P > 0.05). The apoptosis rates at different time point of the HER-2 siRNA III + cisplatin group were all higher than those of the other 2 groups (all P < 0.01). The protein expression levels of the antiapoptotic proteins Bcl-2, survivin, and XIAP were of the HER-2 siRNA III + cisplatin group were significantly lower than those of the cisplatin group, and the protein expression level of the apoptotic protein Smac of the HER-2 siRNA III + cisplatin group significantly higher than that of the cisplatin group (all P < 0.05). CONCLUSIONS: HER-2 siRNA III induces cell apoptosis significantly and increases the sensitivity of the human ovarian carcinoma cells to cisplatin. The mechanism of induction of cell apoptosis by HER-2 siRNA III + cisplatin may be related to the downregulation of Bcl-2, survivin and XIAP protein and the up-regulation of Smac protein.

[Inhibition of invasive and chemotactic abilities of SKOV3 cells by human epithelial growth receptor-2 small interfering RNA].

OBJECTIVE: To investigate the effects of RNA interference (RNAi) targeting human epithelial growth receptor-2 (HER-2) gene on the invasive and chemotactic ability of SKOV3 cells. METHODS: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted (HER-2 siRNAI, HER-2 siRNAII, HER-2 siRNAIII) sequence were selected in the coding region of HER-2 mRNA. Transfection of HER-2 siRNA was conducted with lipofectamine 2000 in ovarian carcinoma cell line SKOV3. The HER-2 gene expression was assessed by real-time PCR and western blot assays. Changes of invasive and chemotactic capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal. RESULTS: Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose, the expression of GAPDH protein was decreased. GAPDH protein gray value in control group, different doses (0.5, 1.0, 1.5, 2.0 microg) GAPDH siRNA interference groups were 0.6855 +/- 0.0259, 0.5698 +/- 0.0275, 0.4542 +/- 0.0296, 0.3341 +/- 0.0178 and 0.1816 +/- 0.0180, respectively. There was a significant difference in each group (F = 198.126, P < 0.01). Both HER-2 siRNAII and siRNAIII could inhibit the expression of HER-2 protein. There was a significant difference in inhibition of HER-2 expression between the siRNAIII (0.1562 +/- 0.0067), siRNAII (0.2162 +/- 0.1589) groups and the other groups (F = 69.461, P < 0.01). In comparison of the relative expression levels in each dose (0.5, 1.0, 1.5, 2.0 microg) group, the difference was significant (F = 174.53, P < 0.01). The relative expression of HER-2 mRNA in HER-2 siRNA group after the 1(st), 3(rd), 6(th) interference were 0.0506 +/- 0.0017, 0.0266 +/- 0.0011 and 0.0154 +/- 0.0020, respectively. There was a decreasing trend in expression levels at different times (P < 0.01). The invasion and chemotactic capacity of SKOV3 cells were decreased after transfection of HER-2 siRNA into the cell line (F = 53.707, P < 0.01 vs F = 11.361, P < 0.01). CONCLUSIONS: HER-2 siRNAIII can silence the mRNA expression in a time and dose dependent manner. HER-2 siRNA can inhibit cell invasion and chemotactic capacity of SKOV3 cells. The application of HER-2 siRNA extends the list of available therapeutic modalities in the treatment of human ovarian cancers.

[Specific inhibition of HER-2 expression in ovarian carcinoma cells by siRNA targeting HER-2].

OBJECTIVE: To investigate the effects of RNA interference (RNAi) targeting HER-2 gene on the biological traits of human ovarian carcinoma. METHODS: siRNA specific to HER-2 gene was synthesized according to the sequence in the GenBank. Human ovarian carcinoma cells of the line SKOV-3 were cultured and divided into 3 groups: control group; non-specific group, transfected with non-specific siRNA; and specific group, transfected with specific HER-2 siRNA. On the 5th day after transfection cisplatin was added into the culture fluid. The expression of HER-2 mRNA and the expression of protein both before and after transfection were detected by RT-PCR and Western blotting. The cell apoptosis was assessed by flow cytometry. The chemosensitivity of transfected cells to cisplatin was determined by MTT method. RESULTS: Since the 3rd day after transfection the expression of HER-2 mRNA of the HER-2 siRNA group was suppressed till 10 days later. On the 7th day after transfection the expression rate of HER-2 protein of the HER-2 siRNA group was (25.5 +/- 0.8)%, significantly lower than those of the nonspecific siRNA group and control group, (95.7 +/- 0.8)% and (96.6 +/- 1.2)% respectively (both P < 0.001). On the 9th day after transfection no expression of HER-2 protein was found in the HER-2 siRNA group. The apoptosis rate of SKOV-3 cells increased time-dependently after transfection in the HER-2 siRNA group and reached the peak, (53.2 +/- 1.0)% on the 6th day, significantly higher than those of the non-specific siRNA group and control group, (4.1 +/- 0.3)% and (4.1 +/- 0.3)% respectively (both P < 0.001). After exposure to cisplatin for 24 hours, the survival rate of the HER-2 siRNA group was (58.4 +/- 0.8)%, significantly higher than those of the nonspecific siRNA group and control group, (68.0 +/- 0.6)% and (67.0 +/- 0.3)% respectively (both P < 0.001). CONCLUSION: siRNA targeting HER-2 synthesized in vitro and transfected into human ovarian carcinoma cells effectively suppresses the HER-2 expression, induces cell apoptosis, and increases the sensitivity to cisplatin of the cells. The successful application of HER-2 siRNA extends the list of available therapeutic modalities in the treatment of human ovarian cancer.