RESUMEN
Metastasisassociated lung adenocarcinoma transcript 1 (MALAT1) is a long noncoding RNA (lncRNA) that has an oncogenic role in some types of cancers, uncluding breast cancer (BC). To investigate the role of MALAT1 in human BC progression, we detected MALAT1 expression levels based on tissue samples from 20 BC cases and 20 healthy controls and found MALAT1 expression levels to be significantly high (P<0.05). Then, we knocked down endogenous MALAT1 in MCF7 cells using MALAT1 short hairpin RNA (shRNA). The results revealed that MALAT1 knockdown could significantly inhibit proliferation, migration, and tube formation in vitro. In addition, miR145 expression inversely changed in BC tissue cases. Furthermore, knockdown of endogenous MALAT1 significantly increased miR145 levels in MCF7 cells. This finding indicated an interaction between MALAT1 and miR145. In addition, knockdown of MALAT1 significantly reduced the expression of vascular endothelial growth factor in MCF7 cells. This outcome revealed that MALAT1 promoted angiogenesis in BC, which may be related to the expression of miR145.
Asunto(s)
Neoplasias de la Mama/genética , MicroARNs/genética , Neovascularización Patológica/genética , ARN Largo no Codificante/genética , Neoplasias de la Mama/patología , Movimiento Celular/genética , Proliferación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Técnicas de Silenciamiento del Gen , Humanos , Células MCF-7 , Neovascularización Patológica/patología , ARN Interferente Pequeño/genética , Transducción de Señal/genéticaRESUMEN
Calcium ion fluctuation is closely related to the transformation of cell cycle. However, little is known about the function of L-type calcium channel in mammalian oocyte and embryo development. We thus studied the roles of L-type calcium channel in mouse oocyte meiotic maturation, parthenogenetic activation and early embryonic development. We used the antagonist Amlodipine to block L-type calcium channel. Oocytes or zygotes were cultured to different time points with 0 µM, 10 µM, 30 µM and 50 µM Amlodipine. Then we checked the rate of first polar body extrusion, spindle formation, asymmetric division parthenogenetic activation and early embryo cleavage. The results showed that Amlodipine treatment did not affect germinal vesicle breakdown, but caused disruption of cytoskeleton organization, symmetric division, formation of mature oocytes with a large polar body, or reduced the first polar body extrusion, depending on its concentrations. Amlodipine treatment also resulted in decreased parthenogenetic activation and arrested early embryonic development. Overall, these data suggest that proper function of L-type calcium channel is critical for oocyte maturation, activation, and early embryonic development.
Asunto(s)
Canales de Calcio Tipo L/fisiología , Embrión de Mamíferos/fisiología , Desarrollo Embrionario/fisiología , Oocitos/fisiología , Amlodipino/farmacología , Animales , Calcio/metabolismo , Bloqueadores de los Canales de Calcio/farmacología , Fase de Segmentación del Huevo , Técnicas de Maduración In Vitro de los Oocitos , Meiosis , RatonesRESUMEN
The developmental potential of post-ovulatory oocytes decreases with aging in vivo and in vitro. In this study, we aimed to investigate the effects of a potent antioxidant caffeine on cortical granules (CGs) distribution in mouse oocytes aging in vivo and in vitro. We found that in vivo administration of 150 mg/kg caffeine caused ovulation of some morphologically abnormal oocytes showing premature exocytosis or congregation of CGs, but significantly decreased abnormal distribution of CGs in oocytes aging for 6 h, 12 h and 18 h in vivo compared to those without caffeine treatment. Unexpectedly, supplementation of oocyte culture medium with 10 mmol/L caffeine accelerated CGs release of oocytes and the normal CG distribution rate dramatically decreased from 6 h in oocytes aging in vitro. It appeared that oocytes showed a high degree of abnormal CG distribution by aging for 18 h, and caffeine might delay oocyte CG exocytosis in vivo, but accelerates CG exocytosis in vitro. Our findings may have implications for improving assisted reproduction technologies.
Asunto(s)
Cafeína/farmacología , Senescencia Celular/efectos de los fármacos , Senescencia Celular/fisiología , Gránulos Citoplasmáticos/metabolismo , Exocitosis/efectos de los fármacos , Oocitos/citología , Oocitos/metabolismo , Animales , Cafeína/administración & dosificación , Células Cultivadas , Femenino , Ratones Endogámicos ICR , Oocitos/patología , Ovulación/efectos de los fármacos , Técnicas Reproductivas Asistidas , Factores de TiempoRESUMEN
Wild animals respond differently to nonconsumptive human activity and such variation depends on multiple factors. We explored the behaviors of Pseudois naynaur and recorded the distance of their responses in Suyu Kou National Forest Park, Helan Mountain, Ningxia Hui Autonomous Region. We categorized their behavioural responses as no response, vigilance and flight and recorded the response initiation distance. We compared distances according to disturbance source, group size, group type and sex. Our results showed that Pseudois naynaur showed stronger responses to humans than vehicles. The distance at which the subject of the group was vigilant in small group (group size less than three) was significantly more than that of larger groups (group size more than three). The flight initiation distance in small groups (less than five) was significantly more than bigger groups. The distance of no response behavior did not vary between all male, female or mixed groups. The distance of vigilance behavior when the subject of the group first encountered the disturbance in male groups was significantly greater for female and mix groups, flight initiation distance in female groups was greater than that of mixed groups. In the mixed group, no significant variation on sex was found among all three types of behaviors.
Asunto(s)
Ecosistema , Cabras/fisiología , Actividades Humanas , Animales , China , Femenino , Humanos , MasculinoRESUMEN
5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a new ablation treatment for tumors, while its function mechanism in cervical cancer has not been elucidated. In this study, we investigated the effects of ALA-PDT on the cervical cancer cell line Me180, to search for optimal parameters of PDT. ALA-PDT on proliferation of Me180 was examined by MTT assay to find the optimal function parameters of ALA-PDT. Apoptosis was observed by using AnnexinV-FITC/PI double staining, Hoechst 33342 staining and May-Grünwald-Farbstoff Giemsa staining. Furthermore, we established a tumor model and 6 mice of each group underwent measurement of the tumor size on days 3, 7, 14, and 21 after treatment. The mRNA expression of survivin, bcl-2, p53, bax and bad in Me180 cells were detected by real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) in vitro and in vivo. Finally, we compared the effects between topical and intravenous administration. Based on the above studies, we found ALA-PDT induced apoptosis and G0-G1 phase arrest of Me180 cells. The tumor volume of the topical administration and PDT group was the smallest at 7-14 days post-PDT. H&E staining showed remarkable subcutaneous necrosis in the PDT groups. The mRNA expression of survivin and bcl-2 in Me180 cells were suppressed post-PDT. Topical administration of PDT is recommended in treating cervical cancer so as to minimize the side-effects and inconvenience of phototoxic reaction brought by PDT. Our data may contribute to making the mechanism of PDT on cervical cancer clearer and give some useful suggestions for clinical application.
Asunto(s)
Ácido Aminolevulínico/uso terapéutico , Fotoquimioterapia , Fármacos Fotosensibilizantes/uso terapéutico , Neoplasias del Cuello Uterino/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patologíaRESUMEN
BACKGROUND & OBJECTIVE: 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a new ablation treatment for tumors, while its function mechanism in cervical cancer is unclear. This study was to investigate the effects of ALA-PDT on cervical cancer cell lines. METHODS: The effects of ALA-PDT on proliferation of 8 human cervical cancer cell lines were examined by MTT assay to find out the optimal function parameters of ALA-PDT and the most sensitive cell line. The effect of ALA-PDT on apoptosis of cervical cancer cell line Me180 was investigated by using Annexin V-FITC/PI double staining, Hoechst 33342 staining and May-Grunwald-Farbstoff Giemsa staining. Cell cycle of Me180 cells was observed by flow cytometry with PI staining. The effect of ALA-PDT on the expression of survivin in Me180 cells was detected by Western blot and real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Among the 8 cell lines, Me180 cell line was most sensitive to ALA-PDT; 2 mmol/L ALA, 10 J/cm(2) light dose and 3-hour treatment were the optimal function parameters of ALA-PDT, and the 50% inhibition concentration (IC(50)) of ALA-PDT under this situation for Me180 cells was 7.28x10(-4) mmol/L. ALA-PDT induced apoptosis and G0/G1 phase arrest of Me180 cells, and suppressed the mRNA and protein expression of survivin in Me180 cells. CONCLUSION: ALA-PDT can inhibit the proliferation and induce apoptosis of cervical cancer cell line Me180 in vitro, which may relate with the suppression of survivin expression.
Asunto(s)
Ácido Aminolevulínico/farmacología , Apoptosis/efectos de los fármacos , Proteínas Asociadas a Microtúbulos/metabolismo , Fotoquimioterapia , Neoplasias del Cuello Uterino/patología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Femenino , Humanos , Proteínas Inhibidoras de la Apoptosis , Proteínas Asociadas a Microtúbulos/genética , Fármacos Fotosensibilizantes/farmacología , ARN Mensajero/metabolismo , Survivin , Neoplasias del Cuello Uterino/metabolismoRESUMEN
OBJECTIVE: To study the effects of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on human cervical cancer, and to identify the best administration regimen with the least phototoxic reaction. METHODS: Seventy-two BALB/c nude mice underwent subcutaneous injection of human cervical cancer cells of the line Me180 so as to establish animal models, and then randomly divided into 6 equal groups: pure topical administration group (undergoing injection of ALA 60 mg/kg around the tumor), pure intravenous administration group (undergoing injection of ALA 250 mg/kg into the caudal vein), topical administration and PDT group (undergoing injection of ALA 60 mg/kg around the tumor and radiotherapy of 630 nm He-Ne laser 6 h after the drug administration), intravenous administration and PDT group (undergoing injection of ALA 250 mg/kg into the caudal vein and radiotherapy of 630 nm He-Ne laser 6 h after the drug administration), and control group (undergoing none of the treatment). 24 h later 6 mice from each group were killed with their tumors taken out. On the days 3, 7, 14, and 21 after treatment the remaining mice underwent measurement of the tumor size. HE staining and pathological examination were performed. Immunohistochemical study was conducted to detect the protein expression of the apoptosis-inhibiting genes of survivin and vascular endothelial growth factor (VEGF). RT-PCR was used to detect the mRNA expression of the apoptosis-inhibiting gene of Bcl-2 and apoptosis-promoting genes of P53, Bax, and Bad. RESULTS: The tumor volumes after treatment of the pure topical administration and pure caudal vein administration groups were (0.09 +/- 0. 02) cm3 and (0.14 +/- 0.04) cm3 respectively, both significantly smaller than that of the control group [(0.67 +/- 0.06) cm3, both P < 0.01]. The tumor volume of the pure topical administration group 7-14 days after treatment was the smallest. HE staining showed remarkable subcutaneous necrosis in the 2 PDT groups. Immunohistochemistry showed remarkable down-regulation of protein expression of survivin and VEGF in the PDT groups. RT-PCR showed that the mRNA expression levels of survivin and Bcl-2 were both significantly lower than that of the control group (both P < 0.01), and the mRNA expression levels of P53, Bax, and Bad were higher than those of the control group, however, not significantly (all P > 0.05). CONCLUSION: ALA-PDT is effective in treatment of cervical cancer. Topical ALA administration + PDT is recommended in treating cervical cancer so as to minimize the side-effects and inconvenience of phototoxic reaction brought by PDT.
Asunto(s)
Ácido Aminolevulínico/uso terapéutico , Fotoquimioterapia , Neoplasias del Cuello Uterino/tratamiento farmacológico , Ácido Aminolevulínico/administración & dosificación , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Femenino , Humanos , Inmunohistoquímica , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Proteínas Asociadas a Microtúbulos/biosíntesis , Survivin , Neoplasias del Cuello Uterino/patología , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
OBJECTIVE: To examine and follow up cervical cytology of pregnant and postpartum women and study their cytopathologic characteristics, so as to determine screening and managing programs for abnormal cervical cytology. METHODS: Totally 5296 patients in pregnancy and postpartum, in which 3729 by computer-assisted cytology test and 1567 by liquid-based monolayers cytology test, were examined and diagnosed by the Bethesda System made in 2001. Those proven epithelial abnormalities patients were followed up until the lesions regressed to normal. The remaining patients, who exhibited persistent abnormalities or progression, were given further examinations. RESULTS: The positive rate of cervical cytological test was 8.72% (462/5296), including squamous carcinoma (SCA) 1 case (0.02%), high grade squamous intraepithelial lesion (HSIL) 34 cases (0.64%), low grade squamous intraepithelial lesion (LSIL) 145 cases (2.74%), atypical glandular cells (AGC) 5 cases (0.09%), atypical squamous cells cannot exclude HSIL (ASC-H) 14 cases (0.26%), atypical squamous cells of undetermined significance (ASC-US) 263 cases (4.97%). The 419 proven cytological abnormality cases were followed up successfully. The total transnegative rate in three months was 73.74% (309/419), in which 303 cases (72.32%) persisted normal status for more than six months after regression. And the transnegative rate of ASC-US, ASC-H, AGC, LSIL, and HSIL were 79.56%, 64.29%, 100%, 72.14% and 44.12%, respectively. Forty-six cases received biopsy directed by colposcopy. The distribution of coincidence of cytopathologic and histopathologic diagnosis was: SCA 1 case (100%), HSIL 10 cases (76.92%), LSIL 13 cases (65%), ASC-H 2 cases (50%), ASC-US 3 cases (37.50%), total 29 cases (63.04%). CONCLUSIONS: We should cast more attention to screening cervix lesions in pregnant and postpartum women. Their cytopathologic characteristics are liable to make the clinician give a false positive diagnosis. So we propose to follow up them closely and to lower the indication of biopsy.