Mechanism of resveratrol regulating ovarian cancer cell apoptosis by inhibiting HMGB1 / 中华内分泌外科杂志

Objective:To investigate the effect of resveratrol on apoptosis of ovarian cancer cells and its molecular mechanism, and to find a potential new target for the treatment of ovarian cancer.Methods:Ovarian cancer SKOV-3 cells were divided into control group and resveratrol group.The survival rate of SKOV-3 cells treated with resveratrol was measured by MTT assay. 24 h after resveratrol intervention in SKOV-3 cells, Western blot was used to detect the expression of Bcl-2, Bax, Caspase-3, HMGB1, TLR4 and NF-?B. Ovarian cancer SKOV-3 cells were transfected with si-NC, si-HMGB1, Rb1+pcDNA3.1 or Rb1+pcDNA3.1-HMGB1, and the sensitivity of cells to resveratrol was detected by MTT assay. The transfected cells were treated with resveratrol and apoptosis was detected by flow cytometry.Results:Resveratrol could inhibit the growth of ovarian cancer SKOV-3 cells, and the higher the concentration of resveratrol, the more significant the inhibitory effect. The expression level of HMGB1 in control cells and resveratrol group was 1.24±0.15 and 0.86±0.11, respectively. 25μM resveratrol inhibited HMGB1 protein level ( P<0.01) . The sensitivity to resveratrol was increased after HMGB1 was downregulated, and the apoptosis of SKOV-3 cells was promoted. HMGB1 overexpression increased the resistance to resveratrol and inhibited the apoptosis of SKOV-3 cells. TLR4 expression quantity control and resveratrol cells were 0.98±0.12 and 0.63±0.08, amount of NF-?B expression were 1.21±0.14 and 0.45±0.07, 25 μM resveratrol could cut TLR4 and NF-?B protein levels. Conclusions:Resveratrol can promote apoptosis of ovarian cancer SKOV-3 cells. This mechanism may be related to the down-regulation of HMGB1/TLR4/NF-?B, which provides a basis for resveratrol treatment of ovarian cancer.

Effect of anemoside B4 on proliferation, migration and apoptosis of ovarian cancer SKOV3 cells and its mechanism / 中华内分泌外科杂志

Objective:To investigate the effects of anemoside B4 on proliferation, apoptosis and migration of ovarian cancer SKOV3 cells through a variety of biological methods, and further to explore its mechanism.Methods:Ovarian cancer SKOV3 cells were cultured in vitro. CCK-8 method was used to detect the proliferation of SKOV3 cells treated with different concentrations of anemoside B4, and IC50 value was calculated. Flow cytometry was employed to detect the apoptosis of cells treated with IC50 concentration of anemoside B4 for different time length; Transwell method was used to detect the migration and invasion of cells treated with IC50 concentration of anemoside B4 for different time length. Western blot was used to detect changes in the expression of related proteins.Results:Anemoside B4 can effectively inhibit the proliferation of SKOV3 cells, showing a concentration-dependent IC50 value of 6.08±0.56 μM, and the inhibitory effect is stronger than the positive control drug cisplatin, with statistically significant difference (P<0.05) . Flow cytometry showed that anemoside B4 could induce SKOV3 cells apoptosis, reduce Bcl-xl expression, and up-regulate the expression of Bax, cleaved-caspase-3 and PARP. Compared with the group of 0 h, the difference was statistically significant (P<0.05) . Crocetin could down-regulate the expression of N-cadherin and up-regulate the expression of E-cadherin, thereby inhibiting the migration of SKOV3 cells. Anemoside B4 could inhibit the expression of JAK/STAT3 signaling pathway proteins.Conclusion:Anemoside B4 can effectively inhibit the proliferation of cervical cancer cells, and induce SKOV3 cell apoptosis by regulating the JAK/STAT3 signaling pathway to inhibit their migration. Crocetin has great potential in the research and development of ovarian cancer therapy.

Effect of miR-581 on Autophagy of Ovarian Cancer SKOV3 Cells by Regulating FOXO1 / 肿瘤防治研究

Objective To explore the effect and mechanism of miR-581 on the autophagy of ovarian cancer SKOV3 cells. Methods miR-581 mimics and miR-581 NC were transfected into SKOV3 cells, and the transfection efficiency was detected by qRT-PCR. After successful transfection, Western blot was used to detect autophagy-related proteins expression in SKOV3 cells. TargetScanHuman database predicted miR-581 target genes, and Western blot verified the role of miR-581 and target genes. Results Overexpression of miR-581 significantly inhibited the expression of autophagy-related proteins LC3 Ⅱ and Beclin1 (P < 0.01); miR-581 negatively regulated FOXO1 expression (P < 0.01) and FOXO1 promoted autophagy of SKOV3 cells (P < 0.01). Conclusion miR-581 affects the autophagy of ovarian cancer SKOV3 cells by regulating the expression of FOXO1.

Effect of icaritin on proliferation and apoptosis of human ovarian cancer cells SKOV3 / 中国中药杂志

Based on the PI3K/Akt signaling pathway, this study aimed to observe the proliferation and apoptosis of ovarian cancer SKOV3 cells at different concentrations of icaritin, in order to explore the possible molecular mechanisms. The research object was ovarian cancer SKOV3 cells. The cells were divided into the control group and icaritin groups(5, 10, 20 μmol·L~(-1)), and administrated with drugs for 48 hours. The cell counting kit-8(CCK-8)assay was used to detect the inhibitory effect of icaritin on the proliferation of ovarian cancer SKOV3 cells. The proliferation ability of the SKOV3 cells was detected by EdU assay. Hoechst 33342 fluorescence staining was used to observe the apoptotic morphology of SKOV3 cells in each group. The distribution of cell cycle and the apoptosis rate of each group were detected by flow cytometry. Quantitative Real-time PCR was used to detect mRNA expressions of PTEN, PI3K, Akt in each group of cells. Protein expressions of PTEN, PI3K, Akt and p-Akt were measured by Western blot. The results showed that the cell inhibition rates of icaritin groups were significantly increased compared with the control group(P<0.05). The rates of EdU-positive cells of icaritin groups were significantly decreased(P<0.05). SKOV3 cells in icaritin groups showed morphological changes of apoptosis. Apoptosis rates of icaritin groups were significantly increased(P<0.05). The proportions of cells in G_0/G_1 phase of icaritin groups were decreased(P<0.05), while the proportions of S phase cells were increased(P<0.05). The gene and protein expressions of PTEN in icaritin groups were elevated(P<0.05). The gene expressions of PI3K and Akt in icaritin groups were down-regulated(P<0.05). The protein expression of PI3K and p-Akt in icaritin groups were reduced(P<0.05). These results indicated that icarin may inhibit the proliferation of ovarian cancer SKOV3 cells in vitro, induce cell apoptosis and affect the cycle distribution of cells by inhibiting the PI3K/Akt signaling pathway.

Effects of EGCG on growth and expression of VEGF and PCNA in ovarian cancer xenografts of nude mice / 中国药理学通报

Aim To explore the effects of EGCG on xenografts of ovarian cancer in nude mice and its possible mechanism. Methods Nude mice xenografts of ovarian cancer SK0V3 cells were established and divided into five groups after tumor formation, in which three groups were given EGCG (10, 30, 50 mg • k g-

Effects of rhein derivatives 4 a containing amide structure on migration and invasion in ovarian cancer cells / 中国药理学通报

Aim To investigate the effect of Rhein derivative 4a containing amide structure on migration and invasion in ovarian cancer SKOV3 cells and its possible mechanism. Methods Ovarian cancer SK0V3 cells were used as target cells. Molecular docking and West-em blot were used to detect the regulatory effect of derivative 4a on Racl protein. CCK8, HE staining, Scratch and Transwell assay were used to detect the effects of derivative 4a on the proliferation, morphology , migration and invasion of SK0V3 cells, respectively. Western blot was employed to determine the expression of matrix metalloproteinases and EMT-related proteins. Results Derivative 4a could effectively bind to Racl protein, and the binding energy was-29. 10 kcal • mol"1, which was significantly lower than that of Rhein; it also could down-regulate the expression of Racl protein in SK0V3 cells. Derivative 4a could significantly inhibit the proliferation, invasion and migra tion of SKOV3 cells, and induce a large amount of cellular vacuolation; derivative 4a could also down-regu-\ late the expression of MMP-2 and MMP-9, up-regulate the expression of EMT epithelial marker protein E-ca-derin but down-regulate the expression of vimentin and j3-cantenin. Conclusions Derivative 4 a can inhibit the proliferation, migration and invasion of ovarian cancer SK0V3 cells. The mechanism may relate to its targeted regulation of Racl, thereby inhibiting the secretion of matrix metalloproteinases, up-regulating the expression of key molecule E-caderin and down-regula-ting the expression of Vimentin and (3-cantenin in EMT i process.•.

Effect of silencing sirtuin 3 on resveratrol-induced apoptois of human ovarian cancer SKOV3 cels / 吉林大学学报(医学版)

Objective: To investigate the effect of silencing sirtuin 3 (Sirt3) on the apoptosis of human ovarian cancer SKOV3 cells induced by resveratrol (Res), and to explore its mechanism of promoting apoptosis. Methods: The human ovarian cancer SKOC3 cells were cultured with different concentrations 0, 2. 5, 5. 0, 10.0, 20.0, 40.0 and 80.0 mg · L-1) of Res for 24 h. The survival rate of cells was measured by MTT assay. The SKOV3 cells were randomly divided into control group, Sirt3 inhibitory 3-1H-1, 2, 3-triazol-4-yl) pyridine 3-TYP group, Res group and 3-TYP+Res group. After 24 h of culture, the inhibitory rates of proliferation of the cells in various groups were detected by MTT assay; the nuclei were stained with Hoechst 33342, and the morphorgy nucleus was observed by laser confocal microscope; reactive oxygen species (ROS) probe was used to detect the intracellular ROS levels; Western blotting method was used to detect the expression levels of Sirt3, Bax, Bcl-2 and cleaved caspase-3 proteins in the cells in various groups. Results: The results of MTT assay showed that the survival rates of SKOV3 cells were significantly decreased with the increase of concentration of Res, and the median inhibitory concentration (IC50) was 42. 73 mg · L-1. Compared with control group, the inhibitory rates of proliferation of cells in Res group and 3-TYP+Res group were significantly decreased (P0.05); the protein expression levels of Sirt3 and Bcl-2 proteins in Res group were significantly decreased (P< 0.05), and the expression levels of Bax and cleaved caspase-3 proteins were significantly increased (P<0.05). Compared with Res group, the expression levels of Bax and cleaved caspase-3 proteins in 3-TYP + Res group were significantly increased (P<0.05), and the expression levels of Bcl-2 and Sirt3 proteins in 3-TYP+Res group were significantly decreased (P<0.05). Conclusion: Res can induce the apoptosis of SKOV3 cells, and the inhibition of Sirt3 expression by 3-TYP can enhance the effect of Res.

Identification of High-affinity Novel Targeted Peptides for the Treatment of Human Ovarian Cancer

Background: To investigate the potential of the phage display-identified tumor cellbinding peptide as a biomarker of epithelial ovarian cancer using phage display technology. Method: The Ph.D.-7 Phage Display Peptide Library was used to identify the specific conjugated phages with SKOV3 epithelial ovarian cancer cells, while Chinese hamster ovary cells formed the basis. After employing the rapid differential screening method invitro, the enzyme-linked immunosorbent assay (ELISA), DNA sequencing, immunohistochemistry, immunofluorescence, and the competitive inhibition test of synthetic peptides were used to determine the affinity and specificity of the phages with SKOV3 cells. Results: Using bio panning, we screened the phages, showing a 3590-fold increase after the third round. A total of 61 titers of the phage were randomly selected for ELISA and 10 kinds of the phages with an optical density >0.5 were used for DNA sequencing. Clones of the phage TRRNIPN were derived from DNA sequencing based on ELISA, exhibiting both the brown granular phenomenon and green fluorescence. The specific targeted peptide TRRNIPN was incorporated in tumor cells through the competitive inhibition test. Conclusion: The results of our study indicate that the phage display identified polypeptide TRRNIPN may be an effective biomarker for the early diagnosis and targeted therapy of ovarian cancer

Effects of compound daqiqi decoction combined with cisplatin on inhibition of ovarian cancer skov3 cells / 中国药学杂志

OBJECTIVE: To investigate the inhibitory effect of compound Daqiqi decoction(CDQD) combined with cisplatin on subcutaneously transplanted ovarian cancer in nude mice and its related mechanisms. METHODS: The 60 subcutaneously transplanted model of nude mice was established with human ovarian cancer cell line SKOV3, and divided into 6 groups randomly, each group of 10 nude mice, which were model group that was treated with saline, CDQD low dose group with the CDQD dosage of 15.16 g•kg-1, CDQD medium dose group with the CDQD dosage of 30.33 g•kg-1, CDQD high dose group with the CDQD dosage of 60.66 g•kg-1, cisplatin group with the DDP dosage of 3 mg•kg-1 and combined group that was treated with the CDQD dosage of 30.33 g•kg-1 and the DDP dosage of 3 mg•kg-1. Cisplatin was administered once every 3 d, and the remaining drugs were administered once a day. Then,the tumor-bearing mouse model was given the corresponding drugs for 14 consecutive days, and the tumor volume was measured every 3 d. After the end of treatment, the tumor was removed and weighed. The morphology of the tumor cells were observed by HE staining. The apoptosis of tumor cells was detected by TUNEL method. The mRNA and protein expression of miR939 and STAT3 and VEGFA and EGFR in tumor tissues were detected by real-time fluorescence quantitative PCR and Western-blot. RESULTS: The tumor volume and the tumor weight of the treated groups were all decreased(P<0.01). Compared with the model group, the tumor volume and tumor weight of the combined group were less than those of the other groups(P<0.01), and the apoptosis rate of the combined group was significantly higher than other groups(P<0.01).The expression of miR939 and STAT3 and VEGFA were down-regulated and the expression of EGFR were up-regulated in the treatment groups. Compared with the model group, the expression of MiR-939, STAT3 and VEGFA were down-regulated and the expression of EGFR was increased in the treatment group. MiR-939, STAT3, VEGFA expression in the combined group was the lowest(P<0.05), and the EGFR expression was highest(P<0.05). CONCLUSION: Studies have shown that CDQD can inhibit ovarian cancer subcutaneously transplanted tumor in nude mice, and its mechanism may be related to inhibition of MiR-939/STAT3 pathway activation, down-regulation of VEGFA, and up-regulation of EGFR expression. The inhibitory effect of CDQD on ovarian cancer tissues has a concentration dependence. And the combination of CDQD and DDP can enhance the anti-tumor effect of DDP and reduce the side effects of DDP on tumor-bearing mice.

Influence of cocl2 in cisplatin sensitivity of human ovarian cancer skov3 cells / 吉林大学学报(医学版)

Objective: To observe the effect of CoCl on the cisplatin sensitivity of human ovarian cancer SKOV3 cells, and to clarify the possible mechanism. Methods: The SKOV3 cells were Cultured in vitro and randomly divided into control group, CoCT group, cisplatin (DDP) group and CoCT combined with DDP (combination) group. The cells in CoCL group were Cultured in normal cell medium for 20 h after cultured in 200 pmol • L CoCL for 4 h, the cells in DDP group were cultured in normal cell medium containing 10 mg • L DDP for 24 h, and the cells in combination group were cultured in 10 mg • L DDP for 20 h after cultured in 200 //mol • L CoCl • for 4 h. The survival rates of SKOV3 cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1 a ( HIF-la) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method. Rhod 2-AM fluorescence probe was used to observe the levels of Ca in mitochondria in the cells in various groups. Western blotting method was used to observe the expression levels of cytochrome C (cyto C) cysteinyl aspartasc 3 (caspasc 3) and cleaved cysteinyl aspartasc 3 (cleaved caspase 3). Muse apoptosis assay kit was used to detect the apoptotic rates of cells in various groups. Results: Compared with control group, the survival rate of the cells in CoCI group had no significant change (P> 0.05). and the survival rates of the cells in DDP and combination groups were decreased ( P0. 05) . and the expression levels of cyto C. caspase 3 and cleaved caspase 3 in DDP group were increased significantly ( P < 0.05); comparexl with DDP group, they were lower than those in combination group ( P<0. 05). Comparexl with control group∗ the apoptotic rate of SKOV3 cells in DDP group was increased significantly (P<.0. 05); the apoptotic rate of SKOV3 cells in combination group was lowe'r than that in DDP group (P<0. 05). Conclusion: CoCI can redece the mitochondrial apoptosis of human ovarian cancer SKOV3 cells by inhibiting the DDP-inducexl enhancement of iNOS expression and dccrease the sensitivity of SKOV3 cells to cisplatin.

Influence of CoCl2 in cisplatin sensitivity of human ovarian cancer SKOV3 cells / 吉林大学学报(医学版)

Objective:To observe the effect of CoCl2on the cisplatin sensitivity of human ovarian cancer SKOV3cells, and to clarify the possible mechanism.Methods:The SKOV3cells were cultured in vitro and randomly divided into control group, CoCl2 group, cisplatin (DDP) group and CoCl2 combined with DDP (combination) group.The cells in CoCl2group were cultured in normal cell medium for 20hafter cultured in 200μmol·L-1 CoCl2for 4h, the cells in DDP group were cultured in normal cell medium containing 10mg·L-1 DDP for 24h, and the cells in combination group were cultured in 10mg·L-1 DDP for 20hafter cultured in 200μmol·L-1 CoCl2for 4h.The survival rates of SKOV3cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1α (HIF-1α) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method.Rhod 2-AM fluorescence probe was used to observe the levels of Ca2+in mitochondria in the cells in various groups.Western blotting method was used to observe the expression levels of cytochrome C (cyto C) , cysteinyl aspartase 3 (caspase 3) and cleaved cysteinyl aspartase 3 (cleaved caspase 3) .Muse○R apoptosis assay kit was used to detect the apoptotic rates of cells in various groups.Results:Compared with control group, the survival rate of the cells in CoCl2group had no significant change (P＞0.05) , and the survival rates of the cells in DDP and combination groups were decreased (P＜0.05) ;the survival rate in combination group was higher than that in DDP group (P＜0.05) .Compared with control group, the positive expression intensities of HIF-1αin CoCl2and combination groups were increased (P＜0.05) .Compared with control group, the positive expressions of iNOS in DDP and combination groups were increased (P＜0.05) .The Ca2+levels in the cells in DDP group and combination groups were higher than that in control group (P＜0.05) and the Ca2+level in DDP group was higher than that in combination group (P＜0.05) .Compared with control group, the expression levels of cyto C, caspase 3and cleaved caspase 3proteins in the SKOV3cells in CoCl2group had no significant changes (P＞0.05) , and the expression levels of cyto C, caspase 3and cleaved caspase 3in DDP group were increased significantly (P＜0.05) ;compared with DDP group, they were lower than those in combination group (P＜0.05) .Compared with control group, the apoptotic rate of SKOV3cells in DDP group was increased significantly (P＜0.05) ;the apoptotic rate of SKOV3cells in combination group was lower than that in DDP group (P＜0.05) .Conclusion:CoCl2can redece the mitochondrial apoptosis of human ovarian cancer SKOV3cells by inhibiting the DDP-induced enhancement of iNOS expression and decrease the sensitivity of SKOV3cells to cisplatin.

Induction effect of myricetin on apoptosis of human ovarian cancer SKOV3 cells by promoting DRP1-dependent mitochondrial fission / 吉林大学学报(医学版)

Objective: To observe the apoptosis and mitochondrial fission of human ovarian cancer SKOV3 cells after treated by myricetin and dynamin related protein 1 (DRP1) inhibitor mdivi-1 alone or combined, and to explore the mechanism of myricetin in inducing the apoptosis of SKOV3 cells. Methods: The SKOV3 cells were cultured in vitro and randomly divided into control group, mdivi-1 group, myricetin group and combined group. The cells in mdivi-1 group were treated with 50 μmol · L-1 madivi-1 for 1 h followed by common culture medium for 23 h; the cells in myricetin group were treated with 50 g · L-1 myricetin for 24 h; the cells combined group were treated with 50 jumol · L-1 midiv-1 for 1 h followed by 50 g · L-1 myricetin for 23 h. The survival rates of cells in various groups were detected by MTT assay. The apoptoic rates of cells in various groups were detected by Muse14 apoptosis detection kit. The expression levels of Cyt C, caspase3, DRP1 and FIS1 were observed by Western blotting method. The mitochondrial fission of cells in various groups was observed with MitoTracker® Red. Results; Compared with control group, the survival rate of cells in myricetin group was decreased significantly (P<0. 05); compared with myricetin group, the survival rate of cells in combined group was increased significantly (P<0. 05). Compared with control group, the apoptotic rate of cells in myricetin group was increased (P<0. 05); compared with myricetin group, the apoptotic rate of cells in combined group was decreased (P<0. 05). Compared with control group, the expression levels of Cyt C and caspase3 proteins in the cells in myricetin group were increased (P<0. 05); compared with myricetin group, the expression levels of Cyto C and caspase3 proteins in the cells in combined group were decreased (P<0. 05). Compared with control group, the degree of mitochondrial fission of the cells in myricetin group was increased; compared myricetin group, the degree of mitochondrial fission of the cells in combined group was decreased. Compared with control group, the expression levels of DRP1 and FIS1 proteins in the cells in myricetin group were increased (P<0. 05); compared with myricetin group, the expression levels of DRP1 and FIS1 proteins in the cells in combined group were decreased (P<0. 05). Conclusion: Myricetin can induce the apoptosis of human ovarian cancer SKOV3 cells by promoting the DRPl-dependent mitochondrial fission.

Inhibition of C2CD3 regulates the proliferation, invasion and migration of epithelial ovarian cancer through inhibiting Hedgehog pathway / 上海交通大学学报（医学版）

Objective • To explore the relationship between the expression of C2 calcium dependent domain containing protein 3 (C2CD3) in ovarian cancer and clinicopathological parameters, and its effect on the proliferation and invasion of ovarian cancer SKOV3 cells and possible mechanisms. Methods • The expression of C2CD3 protein in ovarian tissue was detected by immunohistochemistry. The proliferation, migration and invasion abilities of SKOV3 cells were detected by EdU assay, wound healing assay and Transwell assay respectively. Western blotting was performed to investigate the expression of C2CD3, Shh, Ptch1, Smo and Gli1 proteins. Results • C2CD3 protein was located in cytoplasm in ovarian cancer. C2CD3 was highly expressed in ovarian cancer compared to normal ovarian tissue. C2CD3 immunostaining was significantly higher in tumor samples in advanced stages (stage III / ) than in early stages (stage / Ⅱ). The staining intensity was significantly correlated with the grade (grade3 vs. grade1+2). The association between C2CD3 expression and age (or tumor type) was not significant. Inhibition of C2CD3 gene significantly weakened the proliferation, invasion and migration abilities of SKOV3 cells, and the expression of Shh, Ptch1, Smo and Gli1 proteins was significantly decreased. Conclusion • The expression of C2CD3 protein is increased in ovarian cancer tissues. Inhibition of C2CD3 gene weakens the proliferation, invasion and migration capacities of ovarian cancer SKOV3 cells, which may be related to the inhibition of Shh, Ptch1, Smo and Gli1 proteins in Hedgehog pathway.

Research of berberine to proliferation and apoptotic mechanism of human ovarian cancer cell line SKOV3 / 中国免疫学杂志

Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell (SKOV3). Methods:Cell proliferation was detected by MTT method. The cell apoptosis was detected by FCM Annexin V/PI double staining and transmission electron microscopy. The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR. The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR. Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0. 05) in dose-and time-de-pendent manner. When combined with cisplatin, berberine showed synergistic anticancer effects. Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene. In addition, berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA. Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.

Research of berberine to proliferation and apoptotic mechanism of human ovarian cancer cell line SKOV3 / 中国免疫学杂志

Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell (SKOV3). Methods:Cell proliferation was detected by MTT method. The cell apoptosis was detected by FCM Annexin V/PI double staining and transmission electron microscopy. The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR. The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR. Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0. 05) in dose-and time-de-pendent manner. When combined with cisplatin, berberine showed synergistic anticancer effects. Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene. In addition, berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA. Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.

Inhititory effect of proliferation and promotion effect of apoptosis of lactacystin combined with carboplatin on ovarian cancer SKOV3 cells / 吉林大学学报(医学版)

Objective:To study the influence of proteasome inhibitor lactacystin (LAC) and carboplatin in proliferation and apoptosis of the human ovarian cancer SKOV3 cells in vitro ,and to clarify the mechanisms. Methods:The SKOV3 ovarian cancer cells were cultured in vitro ;0,2.5,5.0,10.0 and 20.0 μmol· L-1 LAC were used to intervent the SKOV3 cells for 48 h;5 μmol·L-1 LAC was used to intervent the SKOV3 cells for 0, 24,48,and 72 h;the SKOV3 cells were divided into control group (treated without medical intervention),LAC group (treated with 5 μmol · L-1 LAC), carboplatin group (treated with 10, 20, 40 and 80 μmol · L-1 carboplatin),LAC and carboplatin group (treated with 5 μmol· L-1 LAC and 10,20,40,and 80 μmol· L-1 carboplatin,respectively).MTT method and FCM were used to detect the inhibitory rates of proliferation and apoptotic rates of the SKOV3 cells in various groups.Results:The MTT test results showed that the proliferation of the SKOV3 cells were inhibited with the prolongation of time and increasing of LAC concentration;the half inhibitory concentration (IC50 )of LAC at 48 h was 5.36 μmol · L-1 ;compared with carboplatin group,the inhibitory rates of proliferation of SKOV3 cells in LAC and carboplatin groups were significantly increased (P <0.05).The IC50 of carboplatin was dropped from 58.08 μmol·L-1 to 18.37 μmol·L-1 .The FCM results showed that with the prolongation of treated time of LAC,the apoptotic rates of SKOV3 cells were increased;compared with carboplatin group and LAC group,the apoptotic rate of cells in LAC and carboplatin group was increased (P <0.05).Conclusion:LAC can inhibit the proliferation of the ovarian cancer SKOV3 cells and induce the apoptosis, and LAC can enhance the inhibitory effect of proliferation of carboplatin on the ovarian cancer SKOV3 cells.

Induction effect of myricetin on autophagy in SKOV3 cells and promoting effect on mitochondrial fission / 吉林大学学报(医学版)

Objective:To observe the effects of myricetin on autophagy and mitochondrial fission in the human ovarian cancer SKOV3 cells,and to explore its induction effect on autophagy and promoting effect on mitochondrial fission.Methods: The SKOV3 cells were cultured in vitro.0,20,40,and 60 g·L-1 myricetin were used in control group and low, middle, high doses of myricetin groups for 12 h.The changes of mitochondrial membrane potential were detected by flow cytometry;the morphology of mitochondria was observed by MitoTracker Red;the expression levels of dynamin related protein1 (Drp1) and fission 1 (Fis1) in various groups were detected by Western blotting method;and the expression levels of autophagy related protein LC3 were detected by both Western blotting method and immunofluorescence method.Results:Compared with control group, the ratios of decreased mitochondria in different doses of myricetin groups were significantly increased(t=3.27, t=6.85, t=5.49,P<0.05).Compared with control group, the numbers of mitochondria in different doses of myricetin groups were increased, and the mitochondria looked more like gravel.Compared with control group,the expression levels of Drp1 in different doses of myricetin groups were significantly increased (t=4.35, t=3.28, t=6.17,P<0.05), and the expression levels of Fis1 were increased also(t=8.32, t=6.74, t=9.27).The immunofluorescence results showed that the expression levels of LC3 in different doses of myricetin groups were significantly increased with the increase of myricetin dose compared with control group.The Western blotting results showed that the ratios of LC3-Ⅱ/LC3-Ⅰ in middle and high doses of myricetin groups were significantly increased compared with control group(t=3.28, t=4.21,P<0.05).Conclusion: Myricetin can induce the autophagy of SKOV3 cells, and it can also promote the mitochondrial fission.

Inhibitory effect ofγδT cells on proliferation of ovarian cancer SKOV3 cells / 吉林大学学报(医学版)

Objective:To study the inhibitory effect of γδT cells on the proliferation of ovarian cancer SKOV3 cells,and to clarify its possible mechanism of inducing apoptosis. Methods:The human ovarian cancer SKOV3 cells cultured in vitro were used as control group,and theγδT and SKOV3 cells were co-cultured for 72 h as γδT cells treatment group.Laser scanning confocal microscope was used to obeserve the morphological changes of nucleus SKOV3 cells,and the inhibitory rate of proliferation of SKOV3 cells in two groups were detected by MTT method;Transwell Chambers was used to detect the cell migration ability,then the apoptotic rates of SKOV3 cells were tested by flow cytometry (FCM).Results:The apoptotic morphology of nucleus of SKOV3 cells in γδT cells treatment group were found under microscope,such as nuclear shrinkage.The MTT resultes displayed that the inhibitory rate of proliferation of SKOV3 cells in γδT cells treatment group was higher than that in control group (P <0.05).The Transwell Chambers results showed that the number of transmembrane cells in γδT cells treatment group was lower than that in control group,and the migration rate was decreased compared with control group (P <0.05).The FCM results showed that the apoptotic rate of SKOV3 cells in γδT cells treatment group was higher than that in control group (P < 0.05 ).Conclusion:γδT cells can inhibit the proliferation and the migration abilities of ovarian cancer SKOV3 cells,and promote the apoptosis.

Optimal concentration of superparamagnetic iron oxide-short hairpin RNA dual functional molecular probe transfected into ovarian cancer cells in vitro / 北京大学学报(医学版)

Objective:To explore the effects of superparamagnetic iron oxide-short hairpin RNA ( SPIO-ShRNA) dual functional molecular probes of different concentrations on morphology and biological beha -vior of ovarian cancer SKOV3 cells in vitro.Methods:The dual functional molecular probes at an iron concentration of 5, 15, 30, 45, 75, and 100 mg/L were transfected into SKOV3 cells.The transfection rate of the probe was observed by fluorescence microscope .The distribution and content of iron particles in SKOV3 cells were determined by Prussian blue staining , atomic adsorption spectrometer and electron microscopy .Cell viability was observed by cell counting kit-8 ( CCK-8 ) .The apoptosis was detected by flow cytometry .The expression of protein within the cells was detected by Western blot .The changes of the signal intensity were measured by magnetic resonance imaging (MRI).Results: The SPIO-ShRNA dual functional molecular probe was uptaken in aconcentration-dependence manner within a certain range (5-30 mg/L) .When the concentration of the probe was 45 mg/L, the labeling rate of the cell was close to 100%;With the increase of the concentration of probe , the cell survival rate decreased gradual-ly.The cell survival rate of each experimental group were 94.626%±1.050%, 93.373%±1.180%, 91.700%±3.122%, 75.100%±4.362%, 72.983%±3.233%, 71.010%±2.910%,5, 15, 30mg/L cell survival rate was not significantly decreased , the difference was not statistically significant (P=0.226, P=0.068, P=0.475);When the concentration of the probe was greater than or equal to 45 mg/L,the survival rate decreased obviously ( P<0.001);Group of 45 mg/L protein expression rate was 68.905%± 3.510%, When the concentration of the probe was greater than or equal to 45 mg/L, the inhibition rate of the protein expression level of epidermal growth factor receptor was obviously higher than those of 5, 15, and 30 mg/L groups, the difference was statistically significant (P<0.001, P=0.001, P=0.003, all P<0.01);the MRI displayed that the signal intensity was decreased with increasing concentrations of the probe.The signal intensity of 45 mg/L group was 165.55 ±4.92, compared with the blank control group (same volume of phosphate buffer saline ), normal group(unlabeled ovarian cancer SKOV3 cells), 5, 15, and 30 mg/L groups , the signal intensity of 45 mg/L group decreased significantly (all P<0.001).Con-clusion:The dual functional molecular probe can effectively transfect and specifically inhibit the expression of SKOV3 cell lines at the iron concentration of 45 mg/L, and can also be detected by MRI .The role of diagnosis and treatment of the dual functional molecular probe has been initially confirmed .

Grape seed proanthocyanidin extract inhibits proliferation and promotes apoptosis of ovary cancer cell line SKOV3 / 第三军医大学学报

Objective To study the biological effects of grape seed proanthocyanidin extract (GSPE) on human ovary cancer cells and to explore the molecular mechanism. Methods Human ovary cancer cell line SKOV3 were co-cultured with GSPE solution of the terminal concentrations of 25,50,100,200,400 ?g/ml respectively in 96-well plate. At 24,48,72 h after coculture,the following parameters were detected: cell growth curve,the inhibition rate of SKOV3 cells by MTT assay,DNA cycle by FCM,the apoptosis of SKOV3 by TUNEL and Annexin-V labeling method,the mRNA and protein expressions of survivin by semi-quantitative RT-PCR and Western blotting,respectively. Results GSPE dose-dependently inhibited the proliferation of the SKOV3 cells. Treated by GSPE,the progress of SKOV3 cells at S stage into G/M stage was inhibited. TUNEL showed that treated by 25 ?g/ml GSPE for 24,48 h,the apoptotic rates of SKOV3 cells were 31.98%,45.78% respectively. Annexin-V showed that after incubation with 25 ?g/ml GSPE for 24 h,the apoptotic rate was 14.68%. The survivin mRNA and protein expressions were both down-regulated. Conclusion GSPE inhibits the proliferation of malignant human ovary cancer cells and induces their apoptosis. Expression of survivin mRNA and protein may be related to cell growth inhibition and to the apoptosis mediated by GSPE in vitro.