MicroRNA-181 inhibits proliferation and promotes apoptosis of chondrocytes in osteoarthritis by targeting PTEN

2017 ◽  
Vol 95 (3) ◽  
pp. 437-444 ◽  
Author(s):  
Xiao-Feng Wu ◽  
Zi-Hui Zhou ◽  
Jian Zou
Keyword(s):  
Cell Proliferation ◽  
Caspase 3 ◽  
Gelatin Zymography ◽  
Negative Control ◽  
The Other ◽  
Logarithmic Growth Phase ◽  
Proliferation And Apoptosis ◽  
Protein Expressions ◽  
Pten Mrna ◽  
Inhibit Cell Proliferation

Objective: To investigate the effects of microRNA-181 (miR-181) on the proliferation and apoptosis of chondrocytes in osteoarthritis (OA) by targeting PTEN. Methods: The chondrocytes in logarithmic growth phase were selected and divided into 6 test groups: the normal, blank, negative control, miR-181 mimic, miR-181 inhibitor, and miR-181 inhibitor + PTEN-siRNA groups. Reverse transcription qPCR was used to detect the expressions of miR-181 and PTEN mRNA. MTT assay and flow cytometry were performed to detect cell proliferation and apoptosis. The protein expressions of PARP and caspase-3 and the activity of MMP-2 and MMP-9 were detected by Western blotting and gelatin zymography assay. Results: The miR-181 mimic group showed increased miR-181 expression and decreased PTEN expression compared with the other 5 groups. Also, by comparison with the other 5 groups, the cell proliferation rate declined and the rate of cell apoptosis was elevated in the miR-181 mimic group. The MiR-181 mimic group showed remarkably increased protein expression of caspase-3 and PARP compared with the other 5 groups. The activity of MMP-2 and MMP-9 was higher in the miR-181 mimic group than the other 5 groups. Conclusion: MiR-181 could up-regulate the expressions of caspase-3, PARP, MMP-2, and MMP-9, and thereby inhibit cell proliferation and promote apoptosis of chondrocytes in OA by targeting PTEN.

Effects of Papaya Leaf Extract (Carica papaya L.) on Cellular Proliferation and Apoptosis in Cervical Cancer Mice Model

2019 ◽  
Vol 17 (5) ◽  
pp. 265-275
Author(s):  
Y. Peristiowati ◽  
Y. Puspitasari ◽  
Indasah
Keyword(s):  
Cervical Cancer ◽  
Cell Proliferation ◽  
Hela Cells ◽  
Leaf Extract ◽  
Caspase 3 ◽  
Methanol Extract ◽  
Negative Control ◽  
Proliferation Index ◽  
Control Group ◽  
P53 Expression

This study is aimed at analyzing the anticancer properties of papaya leaf extract, specifically the inhibition of cell proliferation and apoptotic induction through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and p53 pathways. Twenty-five mice (Mus musculus), aged 2 months and weighing 20–30 g, was injected with 0.5 mg dexamethasone for 7 days. The mice were then injected intracutaneously with 1 ml of HeLa cells (8 × 106 HeLa cells/microliter). The mice were divided into five groups (5 each): negative control (P1) (5% CMC-Na, sodium carboxymethyl cellulose), treatment II (225 mg/kg BW (body weight) papaya leaves methanol extract), treatment III (450 mg/kg BW), treatment IV (750 mg/kg BW), and treatment PV (2 mg alcohol anticancer drug). Papaya leaf extract treatments were applied for 2 weeks. Then, the tumor tissue was isolated for hematoxylin and eosin staining. Immunohistochemical imaging was used to detect Ki-67, caspase-3, NF-κB, and p53 expression. Further analysis was undertaken using the ImmunoRatio software program. The results indicated that administration of papaya leaf methanol extract significantly increased the expression of NF-κB and p53 at a dose of 450 mg/kg BW. Our results also showed that the mice treated with 450 mg of papaya leaf extract per kg of BW (P3) had the largest increase of caspase-3 expression compared to the negative control group. Papaya leaf ethanol extract decreased the cancer cell proliferation index and increased apoptosis of cancer cells in animal models of cervical cancer; it may also work to increase NF-kB expression and expression of the p53 gene.

scholarly journals Silencing long non-coding RNA ROR improves sensitivity of non-small-cell lung cancer to cisplatin resistance by inhibiting PI3K/Akt/mTOR signaling pathway

Tumor Biology ◽  
2017 ◽  
Vol 39 (5) ◽  
pp. 101042831769756 ◽  
Author(s):  
Hui Shi ◽  
Jin Pu ◽  
Xiao-Li Zhou ◽  
Yun-Ye Ning ◽  
Chong Bai
Keyword(s):  
Cell Proliferation ◽  
Signaling Pathway ◽  
Negative Control ◽  
Mtor Signaling ◽  
Mtor Signaling Pathway ◽  
Control Groups ◽  
Over Expression ◽  
Non Coding Rna ◽  
Protein Expressions ◽  
Long Non Coding Rna

This study aimed to investigate the effects of long non-coding RNA ROR (regulator of reprogramming) on cisplatin (DDP) resistance in patients with non-small-cell lung cancer by regulating PI3K/Akt/mTOR signaling pathway. Human cisplatin-resistant A549/DDP cell lines were selected and divided into control group, negative control group, si-ROR group, ROR over-expression group, Wortmannin group, and ROR over-expression + Wortmannin group. MTT assay was used to determine the optimum inhibitory concentration of DDP. Quantitative real-time polymerase chain reaction and western blotting were applied to detect expressions of long non-coding RNA ROR, PI3K, Akt, and mTOR. Colony-forming assay, scratch test, Transwell assay, and flow cytometry were conducted to detect cell proliferation, migration, invasion, and apoptosis, respectively. Tumor-formation assay was performed to detect the growth of transplanted tumors. Long non-coding RNA ROR expression was high in human A549/DDP cell lines. Compared with the control and negative control groups, the mRNA and protein expressions of PI3K, Akt, mTOR, and bcl-2 decreased, whereas the mRNA and protein expression of bax and the sensitivity of cells to DDP significantly increased. Cell proliferation, migration, and invasion abilities decreased in the si-ROR and Wortmannin groups. In comparison with control and negative control groups, the mRNA and protein expressions of PI3K, Akt, mTOR, and bcl-2 increased, whereas the mRNA and protein expressions of bax decreased, the sensitivity of cells to DDP significantly increased, and cell proliferation, migration, and invasion abilities decreased in the ROR over-expression group. For nude mice in tumor-formation assay, compared with control and negative control groups, the tumor weight was found to be lighter (1.03 ± 0.15) g, the protein expressions of PI3K, Akt, mTOR, and bcl-2 decreased, and the protein expression of bax increased in the si-ROR group. Long non-coding RNA ROR may affect the sensitivity of lung adenocarcinoma cells to DDP by targeting PI3K/Akt/mTOR signaling pathway.

Effect of SRY-Related High Mobility Group Box 9 on Extracellular Signal-Regulated Kinase/p38 Signaling Pathway in Glioma

2021 ◽  
Vol 11 (1) ◽  
pp. 171-175
Author(s):  
Tianlong Quan ◽  
Chunhua Zhang ◽  
Xin Song ◽  
Lu Wang
Keyword(s):  
Cell Proliferation ◽  
Signaling Pathway ◽  
Cell Invasion ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
High Mobility ◽  
Negative Control ◽  
Glioma Cell Line ◽  
High Mobility Group ◽  
Control Group

As a common malignant tumor in neurosurgery, glioma is characterized as high incidence rate, easy to invade, metastasize and recurrent. It is difficult to treat and has a poor prognosis. The gliomas pathogenesis is complex and has not been fully resolved. Therefore, finding effective molecular targets for glioma is beneficial to improve therapeutic effect. The SRY-related high mobility group box 9 (SOX9) gene involves in mammalian development and is significantly increased in glioma. However, SOX9’s role in gliomas is unclear. The glioma cell line U87 was assigned into control group, scramble group that was transfected with siRNA negative control, and SOX9 siRNA group that was transfected with SOX9 siRNA followed by analysis of SOX9 mRNA and protein level by qPCR and Western blot, cell proliferation by MTT assay, cell apoptosis by Caspase 3 activity assay, cell invasion by Transwell assay, and MMP-9 level by ELISA. SOX9 siRNA transfection significantly downregulated SOX9 mRNA and protein expressions, inhibited U87 cell proliferation, enhanced Caspase 3 activity, suppressed cell invasion of U87, decreased the secretion of MMP-9 in the supernatant, and reduced ERK1/2 and P38 phosphorylation levels (P < 0.05). SOX9 can regulate the progression of glioma by regulating ERK/P38 signaling pathway, promoting cell apoptosis, inhibiting cell proliferation, and restraining cell invasion.

Combining Bevacizumab with knocked-down β-catenin reduces VEGF-A and Slug mRNA in HepG2 but not in Caco-2 cell lines

2021 ◽  
Vol 21 ◽  
Author(s):  
Reem Mebed ◽  
Yasser BM Ali ◽  
Nahla Shehata ◽  
Nadia El-Guendy ◽  
Nahla Gamal ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Target Genes ◽  
Cultured Cells ◽  
Mrna Level ◽  
Negative Control ◽  
The Other ◽  
Signalling Pathways ◽  
Mrna Levels ◽  
Protein Expressions ◽  
The Difference

Background: Bevacizumab (Bev) resistance is hypothesized to be overcome by combination with inhibitors of other signalling pathways. Objective: We aimed to study the effect of combining Bev with knocked down β-catenin (Bev-β-cat-siRNA) on the expression of VEGF-A, Slug, NFКB and its two target genes c-Flip and FasR in HepG2. Expression of VEGF-A and Slug was also studied in Caco-2 cells. Methods: Cultured cells were divided into six groups 1) cells treated with Bev only 2) cells treated with β-catenin-siRNA 3) cells treated with Bev-β-cat-siRNA 4) cells treated with negative control 5) cells treated with Bev-negative control and untreated cells. Expressions were assessed using qPCR and western blotting. Results: Bev-β-cat-siRNA significantly reduced the mRNA level of VEGF-A, which was initially increased in response to Bev alone in HepG2 but not in Caco-2. Additionally, Bev-β-cat-siRNA significantly decreased Slug mRNA level compared to Bev only treated HepG2 cells. In contrast, VEGF-A and Slug mRNA levels in Bev only group were remarkably lower than Bev-β-cat-siRNA in Caco-2 cells. Distinct β-catenin and Slug protein expressions were noticed in HepG2 and Caco-2 cells. On the other hand, Bev-β-cat-siRNA remarkably reduced the level of NFКB, FasR and c-Flip compared to Bev only treated HepG2 cells although the difference was not statistically significant. Conclusion: We conclude that, combining Bevacizumab with knocked down β-catenin reduce the expression of VEGF-A and Slug in HepG2 but not in Caco-2 cells.

scholarly journals Garcinol Alone and in Combination With Cisplatin Affect Cellular Behavior and PI3K/AKT Protein Phosphorylation in Human Ovarian Cancer Cells

Dose-Response ◽  
2020 ◽  
Vol 18 (2) ◽  
pp. 155932582092673
Author(s):  
Jie Zhang ◽  
Huan Fang ◽  
Jinguo Zhang ◽  
Wencai Guan ◽  
Guoxiong Xu
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Caspase 3 ◽  
Anticancer Agent ◽  
Nuclear Factor Κb ◽  
Ovarian Cancer Cells ◽  
Cellular Behavior ◽  
Proliferation And Apoptosis ◽  
Induced Apoptosis ◽  
Physiological Benefits

Garcinol is a plant-derived compound that has some physiological benefits to human cells. However, the effect of garcinol on ovarian cancer (OC) cell proliferation and apoptosis is unknown. The current study aimed to examine the effects of garcinol alone and in combination with cisplatin (DDP) on cellular behavior and to explore the expression pattern of PI3K/AKT and nuclear factor-κB (NF-κB) in human OC cells. We found that OVCAR-3 cell viability was decreased after garcinol treatment. Garcinol alone and in combination with DDP significantly inhibited cell proliferation and had a synergistic effect evaluated by CompuSyn software. The cell cycle analysis showed the S phase arrest by garcinol. Furthermore, garcinol alone and in combination with DDP promoted cell apoptosis. The garcinol-induced apoptosis was further confirmed by the detection of cleavage forms of PARP and caspase 3. An increase in proapoptotic factor Bax expression was also found in garcinol-treated cells. Moreover, garcinol significantly decreased the phosphorylation of PI3K and AKT proteins and downregulated the expression of NF-κB. Thus, our data demonstrated that garcinol has the potential to be used as an anticancer agent and may synergize the effect of DDP. These actions are most likely through the regulation of the PI3K/AKT and NF-κB pathways.

scholarly journals Effects of bFGF on the Modulation of Apoptosis in Gingival Fibroblasts with Different Host Ages

2013 ◽  
Vol 2013 ◽  
pp. 1-7
Author(s):  
Kotaro Tanimoto ◽  
Satoru Ohkuma ◽  
Yuki Tanne ◽  
Ryo Kunimatsu ◽  
Naoto Hirose ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Caspase 3 ◽  
Type I Collagen ◽  
Pcr Analysis ◽  
Type I ◽  
Gingival Fibroblasts ◽  
Cultured Fibroblasts ◽  
Proliferation And Apoptosis ◽  
Young Subjects

The purpose of this study was to investigate the effects of basic fibroblast growth factor (bFGF) treatment on the proliferation and apoptosis of cultured gingival fibroblasts (GFs). Human GFs were isolated from the palatal gingival tissues of 16 healthy volunteers ranging in the age from 9 to 35 years old. Cultured GFs were subjected to the analyses for cell proliferation by ELISA assay, gene expression by RT-PCR analysis, and apoptosis potency by caspase-3 assay. The cell proliferation activity and gene expression of type-I collagen and caspase-3 activity were enhanced significantly by the treatment with bFGF in cultured GFs. Furthermore, the activity of caspase-3 in cultured GFs from young subjects was significantly higher than that in GFs from adults. It is shown that bFGF significantly enhances the gene expression of type-I collagen in cultured fibroblasts from human gingival tissues. It also demonstrated that bFGF modulates the apoptosis of periodontal fibroblasts, and the effect is higher in young subjects, indicating a significant role of bFGF in the prevention of scar formation during wound healing.

Expression and role of miR-637 before and after lymphadenectomy for thyroid carcinoma

2021 ◽  
Vol 11 (7) ◽  
pp. 1245-1253
Author(s):  
Yanqing Qu ◽  
Xiaoyu Chu ◽  
Cuihong Dong ◽  
Weijiao Wang ◽  
Xiaojian Zhang
Keyword(s):  
Cell Proliferation ◽  
Thyroid Carcinoma ◽  
Biological Significance ◽  
Negative Control ◽  
Luciferase Reporter ◽  
Proliferation And Apoptosis ◽  
Before And After ◽  
Proliferation And Invasion

Thyroid carcinoma (TC) is a common endocrine malignancy that can be partially relieved by surgery, but its recurrence rate remains high. It is speculated that miR-637 exerts certain influence in its occurrence and development. Accordingly, we included 87 TC patients and 72 concurrent healthy controls as the research participants and purchased human papillary thyroid carcinoma cells with which to study and analyze the biological significance of miR-637. The determination of miR-637 and SH2B1 in peripheral blood and tissues was performed using nanoparticle-assisted polymerase chain reaction assay, and the identification of cell proliferation and apoptosis was made by MTT, Transwell, and flow cytometry. The results indicated that after transfection of miR-637 into TPC-1, the cell proliferation and invasion capacities in the mimics-miR-637 group were significantly reduced as compared to that of the inhibition-miR-637 and negative control (NC)-miR groups (P < 0.05). While transfection of SH2B1 into TPC-1 cells led to significantly enhanced cell proliferation and invasion capacities in sh-SH2B1 group than in si-SH2B1 and NC groups (P < 0.05). Finally, a double luciferase reporter assay identified enormously inhibited fluorescence activity of SH2B1-WT by mimics-miR-637. According to the experimental results, it is concluded that miR-637 expression was low in TC but increased after lymphadenectomy for TC. Moreover, by targeting SH2B1, miR-637 interferes with TC progression, which carries significant implications for future diagnosis and treatment of the disease.

Mir-29c Expression in Glioma and Its Effects on Tumor Cell Proliferation and Apoptosis

2020 ◽  
Author(s):  
Peiquan HUI ◽  
Yuling WANG ◽  
Bing CHEN ◽  
Zengwu WANG ◽  
Shiqiang QIN
Keyword(s):  
Cell Proliferation ◽  
Glioma Cells ◽  
Negative Control ◽  
Expression Level ◽  
Control Group ◽  
Tumor Differentiation ◽  
Blank Group ◽  
Pathological Type ◽  
Proliferation And Apoptosis ◽  
Experimental Group

Background: To investigate the expression of microRNA-29c (miR-29c) in glioma and its effects on cell proliferation and apoptosis. Methods: A retrospective analysis was performed on 76 glioma patients in People's Hospital of Weifang, Weifang, Shandong, China from May 2013 to June 2017 (experimental group) and 63 healthy subjects in the same period (control group). qRT-PCR was used to detect the miR-29c expression. Changes of serum miR-29c expression level and the correlation of miR-29c of glioma patients with the degree of tumor differentiation and pathological type were observed. Cells were grouped before transfection into blank group (no transfection), negative control group (transfected with miRNA NC) and experimental group (transfected with miR-29c mimics). CCK-8 assay was used to detect cell proliferation, flow cytometry to detect apoptosis. Results: Expression of miR-29c in serum was significantly lower in experimental group than that in control group (P<0.05). The expression level of miR-29c of glioma patients increased with the degree of tumor differentiation (P<0.05). miR-29c in serum was not significantly correlated with the pathological type. Conclusion: miR-29c could inhibit the proliferation of glioma cells and promote apoptosis. miR-29c is lowly expressed in glioma, and the overexpression of which in glioma cells can inhibit tumor cells proliferation and promote apoptosis. It may be a tumor suppressor miRNA of glioma, and the expression level of which can be used as reference for evaluating the grade of glioma. It is indicated that the abnormal expression of miR-29c may be a key factor in the occurrence and development of glioma.

Proliferation and apoptosis processes in the seasonal testicular development of the wild Daurian ground squirrel (Citellus dauricus Brandt, 1844)

2017 ◽  
Vol 29 (9) ◽  
pp. 1680 ◽  
Author(s):  
Yingying Han ◽  
Jinqi Zhan ◽  
Ying Xu ◽  
Fengwei Zhang ◽  
Zhengrong Yuan ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Breeding Season ◽  
Caspase 3 ◽  
Nuclear Translocation ◽  
Ground Squirrels ◽  
Nuclear Antigen ◽  
Plasma Testosterone ◽  
Testicular Development ◽  
Cyclin D2 ◽  
Proliferation And Apoptosis

The aim of the present study was to elucidate the regulatory role of cell proliferation and apoptosis in testicular development of wild Daurian ground squirrels during the breeding season (April), the non-breeding season (June) and before hibernation (September). Gross mass and hormonal analysis showed that the testis : body mass ratio and plasma testosterone concentration fluctuated seasonally, with a peak in April and lowest values in June. Similarly, spermatogenesis was fully developed in April but suppressed in June and September. Testicular decellularisation and vacuolisation was seen during the transition from the breeding to the non-breeding season. Furthermore, testicular levels of proliferating cell nuclear antigen, cyclin D2 and caspase-3 protein were significantly increased in June and September. Intriguingly, positive terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling staining and nuclear translocation of caspase-3 in testicular germ cells appeared only during the prehibernation period, whereas accumulation of cyclin D2 in spermatocyte nuclei occurred in September. These findings demonstrate, for the first time, that both cell proliferation and apoptosis are stimulated during the prehibernation period, indicating that a hormonal-regulated balance of testicular germ cell proliferation and apoptosis may play a pivotal role in preparing for testicular recrudescence of wild Daurian ground squirrels.

scholarly journals Midazolam Inhibits the Apoptosis of Astrocytes Induced by Oxygen Glucose Deprivation via Targeting JAK2-STAT3 Signaling Pathway

2015 ◽  
Vol 35 (1) ◽  
pp. 126-136 ◽  
Author(s):  
Li Liu ◽  
Qi You ◽  
Yingfeng Tu ◽  
Quanyi Li ◽  
Lihong Zheng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Caspase 3 ◽  
Glucose Deprivation ◽  
Oxygen Glucose Deprivation ◽  
Control Group ◽  
Protective Effects ◽  
Intrinsic Bioremediation ◽  
Proliferation And Apoptosis ◽  
Astrocytes Proliferation ◽  
Apoptotic Effects

Background: There is an increasing interest in the role of astrocytes contributing to the intrinsic bioremediation of ischemic brain injury. The purpose of this study was to disclose the effects and mechanism of midazolam (MDZ) on the proliferation and apoptosis of astrocytes under oxygen glucose deprivation (OGD) condition. Methods: The astrocytes were assigned randomly into four groups: control group, OGD group, OGD+MDZ group, and OGD+MDZ+IL-6 group. The astrocytes were treated with MDZ at dose of 10 μmol/L in OGD+MDZ group. And in OGD+MDZ+IL-6 group, the astrocytes were treated with MDZ at dose of 10μmol/L and IL-6 at dose of 50 ng/mL. MTT assay was used to assess cell proliferation, and cell apoptosis was analyzed by TUNEL apoptosis assay kit and flow cytometry. Furthermore, the expression of JAK2, p-JAK2, STAT3, p-STAT3, Bcl-2, Bax and Caspase-3 proteins were determined by western blotting assay. Results: Astrocytes proliferation was decreased obviously in OGD group, while MDZ could increase astrocytes proliferation under OGD condition. Moreover, OGD could induce apoptosis in astrocytes and MDZ could play an anti-apoptotic role. However, IL-6, a JAK2 activator, could attenuate cell proliferation and anti-apoptotic effects of MDZ in astrocytes. In addition, the expression of Bcl-2 protein in MDZ group increased markedly, while the JAK2/STAT3 signal proteins, Bax and Caspase-3 proteins decreased relative to OGD group. But IL-6 could counteract the anti-apoptotic effects of MDZ. Conclusion: Midazolam has protective effects on the proliferation and apoptosis of astrocytes via JAK2/STAT3 signal pathway in vitro. We firstly disclose the beneficial roles of midazolam in astrocytes under ischemic condition, which may be a rational treatment selection for ischemic cerebral protection.

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