ABSTRACT
@#[Abstract] Objective: To investigate the effects of silencing monocarboxylate transporter 4 (MCT4) on the proliferation, migration and invasion of prostate cancer PC3 cells and its possible molecular mechanism. Methods: RNA interference technology was used to transfect siRNA-MCT4 (si-MCT4) and negative control plasmid (si-NC) into PC3 cells, respectively. The content of lactic acid in the cell culture medium of transfected PC3 cells was detected by lactic acid assay after culturing for 96 h. The proliferation, migration and invasion ability of PC3 cells were detected by CCK-8 and Transwell assay, respectively. Western blotting was used to detect the silencing effect and the expressions of integrin β 4-FAK-SRC-MEK-ERK signaling pathway associated proteins (integrin β4, p-FAK, p-SRC, p-ERK1/2, p-MEK1/2) and EMT associated proteins (E-cadherin and N-cadherin). Results: PC3 cell line with silenced MCT4 was successfully constructed. Compared with the control group, the content of extracellular lactic acid in the PC3 cell culture medium of the si-MCT4 group was significantly decreased (P<0.01), and the proliferation, migration and invasion of cells were significantly decreased (P<0.05 or P<0.01). Compared with the control group, the protein expressions of integrin β4, p-FAK, p-SRC, p-MEK1/2, p-ERK1/2 and N-cadherin were significantly decreased (all P<0.01), while the protein expression of E-cadherin was significantly increased (P<0.01). Conclusion: Silencing MCT4 can significantly inhibit the proliferation, migration and invasion of PC3 cells, the mechanism of which may be related to the inhibition of lactic acid level in cell culture medium and suppression of integrin β4-FAK-SRC-MEKERK signaling pathway associated proteins as well as EMT associated proteins.
ABSTRACT
@#[Abstract] Objective: To investigate the effect of long non coding RNA (lncRNA) LINC00308 on proliferation, invasion and migration of prostate cancer cells and its related mechanism. Methods: lncRNAs and mRNAs differentially expressed in prostate cancer tissues and adjacent control tissues were screened by gene chip, and LINC00308 and TRIP13 (thyroid hormone receptor interactor13) were identified as the research objects. The effects of LINC00308 on the proliferation, invasion and migration of prostate cancer cells were detected by MTT assay, plate cloning, Transwell and scratch test. The above effects were verified in nude mice xenografts. The effect of LINC00308 on expression of TRIP13 in tumor tissues and cancer cells was detected by Western blotting and immunohistochemistry. Bioinformatics analysis, RIP (RNA immunoprecipitation), qPCR and Double luciferase gene reporter experiments were used to predict and explore the interaction mechanism between miR-361-5p and LINC00308 as well as TRIP13, and plate cloning and Transwell invasion test were used to verify the biological behaviors of cancer cells. Results: Both the microarray results and qPCR confirmed that the expressions of LINC00308 (P<0.01) and TRIP13 (P<0.05) were abnormally high in prostate cancer tissues and four cell lines; cell function test results showed that overexpression of LINC00308 could promote the proliferation, invasion and migration of prostate cancer PC3 cells (all P<0.05), while down-regulation of LINC00308 in prostate cancer cells had the opposite effect. In nude mice. LINC00308 could promote the tumorigenesis of prostate cancer cells in vivo, and increase the expression of TRIP13 both in vivo and in vitro (P<0.05). Bioinformatics analysis, RIP, qPCR and Double luciferase gene reporter results confirmed that miR-361-5p could bind to 3'-UTR of LINC00308 and TRIP13 respectively, and LINC00308 could act as a competing endogenous RNA (ceRNA) by sponging miR-361-5p to regulate the expression of TRIP13. In addition, MTT, plate cloning and Transwell assay confirmed the regulatory interaction among LINC00308 miR-361-5p and TRIP13 from the levels of proliferation, colony formation and invasion in cancer cells. Conclusion: LINC00308, which is abnormally highly expressed in prostate cancer tissues and cells, can inhibit the expression of miR-361-5p and enhance the expression of TRIP13 by exerting its ceRNA function, thus promoting the proliferation, invasion and migration of prostate cancer.
ABSTRACT
OBJECTIVE：To st udy the inhibitory mechanism of pomegranate peel polyphenols （PPP）on the proliferation of human prostate cancer PC 3 cells based on autophagy and apoptosis pathway. METHODS ：CCK-8 assay was used to investigate the effects of PPP with different concentrations （25-300 μg/mL）on PC 3 cell activity after culturing for 24，48，72 h，so as to screen the drug concentration and treatment time. PC 3 cells were divided into control group （complete culture medium ），PPP low- ，medium- and high-concentration groups. After treated for 48 h，flow cytometry and Annexin V-FITC/PI staining were used to detect cell cycle distribution and apoptosis of PC 3 cells. Western blotting assay was used to detect the expression of apoptosis-related protein as Bax ，Bcl-2，as well as the expression of autophagy-related proteins as LC 3，Beclin-1，p62，Atg12 and Atg 16. RESULTS ：The culturing time was chosen as 48 h. IC 50 of PPP was 110 μg/mL，and 50，100，200 μg/mL were chosen as low，medium，high concentrations of PPP. Compared with control group ，the percentage of PC 3 cells at phase G 0/G1 decreased significantly in PPP low- and medium-concentration groups while increased significantly at phase S ；that of PC 3 cells at phase G 0/G1 increased significantly in PPP high-concentration group ； while that of PC 3 cells at phase G 2/M decreased significantly in PPP medium- and high-concentration groups （P＜0.05 or P＜0.01）. The apoptosis rate of PC 3 cells was increased significantly in PPP groups （P＜ 0.05 or P＜0.01）. Compared with control group ，protein expression of anti-apoptosis protein Bcl- 2 and autophagy-related promoting protein p 62 were decreased in PPP groups to different extents ，while protein expression of promoting-apoptosis protein Bax as wells as autophagy-related protein LC 3-Ⅱ/LC3-Ⅰ ration and protein expression of Beclin- 1，Atg5，Atg12 and Atg 16 were increased to different extents ；there was statistical significance in above indexes in PPP high-concentration group and some of above indexes in PPP low- and medium-concentration groups （P＜0.05 or P＜0.01）. CONCLUSIONS ：PPP can inhibit the proliferation of human prostate cancer PC 3 cells，mechanism of which may be related to inducing autophagy and promoting apoptosis.
ABSTRACT
@#[Abstract] Objective: To investigate the effects of exosome originated from bone marrow mesenchymal stem cell (BMSCs) on proliferation, migration and invasion of prostate cancer PC-3 cell and its mechanism. Methods: qPCR was used to detect the expression level of miR-21-5p in prostate cancer cell lines. The morphology of exosomes isolated from BMSCs was observed with an electron microscope. Western blotting was used to detect the expressions of exosome surface markers and the epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin). Dual luciferase reporter gene experiment was used to detect the targeted regulation relationship between miR-21-5p and PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2). PC-3 cells were co-cultured with 10 μl BMSCs exosomes suspension (Exo group), transfected with sh-PHLPP2 or antagomiR, then CCK-8 and Transwell experiments were used to detect changesinproliferation,migrationandinvasionofPC-3cell.Results: miR-21-5p was highly expressed in prostate cancer PC-3 cell line. The exosomes in the supernatant of BMSCs culture fluid were successfully isolated, and the typical vesicle-like structures of exosomes were observed under transmission electron microscope. Exosomes expressed specific proteins such as CD9, CD63 and CD81. In the Exo group, the proliferation, invasion, migration, as well as the expressions of N-cadherin, Vimentin and miR-21-5p in PC-3 cells were significantly higher than those in the control group (all P<0.05). PHLPP2 is a target gene of miR-21-5p. Compared with the control group, the expression of PHLPP2 in PC-3 cells of Exo group and sh-PHLPP2 group was significantly reduced (0.66±0.09, 0.42±0.05 vs 1.09±0.08, all P<0.01); cell viability, invasion and migration were significantly improved (all P<0.01); and E-cadherin expression level was significantly reduced while N-cadherin and Vimentin expressions were significantly increased (both P<0.05). Conclusion: miR-21-5p is highly expressed in prostate cancer PC-3 cell line. BMSC exosome miR-21-5p can increase the proliferation, migration and invasion ability of PC-3 cells through targeted down-regulation of PHLPP2.
ABSTRACT
Objective@#To investigate the effect of small interfering RNA silencing the vitamin D receptor (VDR) on the biological behavior of prostate cancer PC-3 cells.@*METHODS@#We constructed the VDR-shRNA lentiviral vector and determined the mRNA and protein expressions of VDR by RT-PCR and Western blot. Using scratch wound healing and Transwell chamber assays, we detected the changes in the migration and invasiveness of the PC-3 cells after silencing VDR.@*RESULTS@#The VDR-shRNA plasmid significantly interfered the VDR expression and successfully screened the cell lines with stable VDR-shRNA interference. The rate of scratch wound healing was markedly lower in the VDR interference group than in the blank control and LV3 negative control groups (59% vs 73.6% and 77.8%, P 0.05), and so was the count of permeable cells (P 0.05). The migration ability and invasiveness of the VDR-treated cells were remarkably decreased as compared with those of the control cells.@*CONCLUSIONS@#Down-regulated expression of the VDR gene may reduce the migration and invasiveness of prostate cancer cells.
Subject(s)
Humans , Male , Cell Line, Tumor , Cell Movement , Genetics , Cell Proliferation , Down-Regulation , Gene Silencing , Lentivirus , Neoplasm Invasiveness , Genetics , Plasmids , Prostatic Neoplasms , Genetics , Pathology , RNA, Messenger , Metabolism , RNA, Small Interfering , Receptors, Calcitriol , Genetics , Metabolism , Transfection , Wound Healing , GeneticsABSTRACT
Objective To study the effects of S-adenosyl-homocysteine hydrolase inhibitor DZNep (3-Dea zaneplanocin A) on human prostatic cancer PC3 cells growth and further explore its potential value in anti-human prostatic cancer treatment.Methods MTT method was used to analyze the effects of EZH2 gene silencing,EZH2 over-expression and DZNep treatment on cell proliferation.Gene expression of EZH2,Bax and Bcl-2 in PC3 cells was detected with western blot.Results DZNep could significantly inhibit PC3 cell growth and induce apoptosis which was identified with Bax expression up-regulation and Bcl-2 expression down-regulation.EZH2 knock-down inhibited PC3 cells growth,and over-expression of EZH2 partially counteract the inhibitory effects of DZNep on PC3 cells growth.Conclusions DZNep can inhibit PC3 cells growth by targeting EZH2 inhibition which leads to endogenous apoptosis.DZNep has the potential value in the treatment of human prostatic cancer.
ABSTRACT
<p><b>Objective</b>To investigate the relationship of the expression of vasodilator-stimulated phosphoprotein (VASP) with the metastasis and prognosis of prostate cancer.</p><p><b>METHODS</b>Prostate cancer PC3 cells were infected with VASP shRNA and control shRNA lentiviruses, respectively. The invasive ability of the PC3 cells was determined by transwell migration assay, the expression of VASP in the prostate cancer tissue from 56 patients was detected by immunohistochemistry, and the survival rate of the patients was analyzed according to the VASP expression levels and follow-up data after radical prostatectomy.</p><p><b>RESULTS</b>VASP shRNA lentivirus significantly inhibited the expression of VASP and decreased the invasive ability of the PC3 cells as compared with the results obtained in the scramble shRNA and blank control groups (P<0.05). The survival analysis of the 56 prostate cancer patients showed that the time of biochemical recurrence was markedly shorter in the VASP positive and strongly positive groups than in the VASP-negative cases (P<0.05), but with no statistically significant difference between the former two groups (P>0.05).</p><p><b>CONCLUSIONS</b>VASP is involved in the regulation of the invasive ability of prostate cancer PC3 cells, and the differences in the VASP expression are related to the prognosis of prostate cancer.</p>
Subject(s)
Humans , Male , Cell Adhesion Molecules , Metabolism , Cell Line, Tumor , Immunohistochemistry , Lentivirus , Microfilament Proteins , Metabolism , Neoplasm Metastasis , Phosphoproteins , Metabolism , Prognosis , Prostatectomy , Prostatic Neoplasms , Pathology , General Surgery , RNA, Small Interfering , Survival RateABSTRACT
Objective To observe the proliferation inhibition and apoptosis promotion effect of oridonin on PC-3 cells.Methods PC-3 cells were treated with different concentrations of oridonin.MTT assay and drug concentration-time survival curve were used to test the effect of oridonin on the PC-3 cells.The percentage of earlier apoptosis cells was analyzed by flow cytometry.The protein expression of caspase-3,Bcl-2,and Bax in the PC-3 cells was detected by Western blot.Results Oridonin effectively inhibited the proliferation of PC-3 cells in both concentration- and time-dependent manner,and the IC50 of PC-3 cells was 10.29 μmol/L.Hochest33258 staining and flow eytometry deteced that oridonin induced the apoptosis of PC-3 cells in a concentration-dependent manner (P ＜ 0.05 ).Oridonin down-regulated Bcl-2,up-regulated Bax protein,and activated caspase-3 in a concentration-dependent manner in the PC-3 cells.Conclusion The apoptosis of PC-3 cells induced by oridonin might be associated with the mitochondrial pathway.
ABSTRACT
Objective To study the effect of small heterocyclic compound 8-oxo-3-thiomorpholino-8H-acenaphtho pyrrole-9-carbonitrile(S1) on apoptosis of human androgen independent prostatic carcinoma cells line(PC3) and its mechanism.Methods PC3 cells were cultivated,and divided into different groups: 10.00,5.00,1.00,0.50,0.10,0.05 and 0.01 ?mol?L-1 S1 groups,meanwhile,PC3 control group and cyclophosphamide group were set up.MTT was used to detect the inhibitory rate of PC3 cell proliferation.Flow cytometry was used to detect the inducing effect of S1 on apoptosis of PC3 cells.Caspase 3,8,9 kits were used to detect apoptosis route.Results The inhibitory rates of PC3 cells induced by 0.10-10.00 ?mol?L-1 S1 were significantly higher than that in cyclophosphamide group(P
ABSTRACT
0.05).The apoptotic percentage was increased significantly after X-rays irradiation with the dose of 6 Gy(P
ABSTRACT
Objective To investigate the mechanism of Kanglaite combined with 5-Fu in treating pancreatic carcinoma through studying the changes of Bcl-2,Bax and vascular endothelial growth factor(VEGF) in pancreatic carcinoma cell and the relationship between apoptosis and these changes.Methods The model of athymic mouse subcutaneous transplantation tumor of human pancreatic carcinoma PC-3 cell was established;the expression of Bcl-2,Bax and VEGF were detected in malignant cells using immunohistochemistry;the apoptosis proportion was detected by adopting flow cytometry;the tumor size was detected.Results Kanglaite combined with 5-Fu could obviously depress the expression of Bcl-2 in protein level in malignant cells;apoptosis proportion had no effect on Bax expression;Kanglaite could obviously depress the expression of VEGF in malignant cells.The apoptosis proportion was higher and protein expression of Bcl-2 was obviously lower in group of Kanglaite combined with 5-Fu than that in Kanglaite group or 5-Fu group.What was more,the protein expression of VEGF and the tumor volum was lowest in group of Kanglaite combined with 5-Fu.Conclusion The effect of Kanglaite united with 5-Fu on treatment of pancreatic cancer is better than that of Kanglaite or 5-Fu alone.
ABSTRACT
AIM To explore the inhibition in in vitro p roliferation human prostate cancer PC 3 cells induced by melatonin and the effe ct of combination of melatonin and 5-Fu. METHODS Cell growth cu rve, MTT assay, and acid phosphatase(ACP)activity were used to determine cell proliferation. Coomassie brillient blue assay and modified diphenylamine assay w ere used to measure the content of protein and DNA in PC 3 cells. ACP activity was measured by modified phenyl phosphat e method of kind and king and modified method of lowry. Co-administration effec t of MLT and 5-fluorouracil(5-Fu)on PC 3 cell proliferation was determined by MTT assay. RESULTS The IC 50 of MLT for inhibiting PC 3, CoLo205, K562, and HeLa cell proliferation was(1 20?0 30),(1 27?0 12) ,(1 60?0 16)and(2 25?0 11)mmol?L -1 , respectively. And MLT inhib ited all above cells proliferation in a concentration-dependent manner. The protein and DNA content and ACP activity in PC 3 cell treated with 2 mmol?L -1 MLT for 96 h were redu ced 58 77%, 54 97% and 89 24%, respectively. When PC 3 cells were treated wi th MLT in combination with 5-Fu 0 5 mg?L -1 , the inhibition rate enhance d. CONCLUSION MLT inhibited PC 3, CoLo205, K562, and HeLa cell p roliferation, and reduced the content of protein and DNA and ACP activity in PC 3 cell. MLT and 5-Fu co-administration enhanced the inhibition rate of PC 3.er.TheproteinandDNAcontentandACPactivityinPC3 celltreatedwith 2mmol?L-1MLTfor 96hwerereduced5 8 77% ,5 4 97%and 89 2 4% ,respectively .WhenPC3 cellsweretreatedwithMLTincombinationwith5 Fu 0 5mg?L-1,theinhibitionrateenhanced .CONCLUSION MLTinhibitedPC3 ,CoLo2 0 5 ,K5 62 ,
ABSTRACT
Purpose:To study the effect of sodium selenate on the viability and proliferation of PC-3 cell, a kind of human prostate cancer cell. Methods:Sodium selenate was administered to PC-3 cells, and MTT assay and ~(3)H-TdR adulterated assay were used to estimate the viability and proliferation of cell. Results:① When cells were treated with Sodium selenate for 24 h, the optical density (A) of middle-dose group decreased significantly, and the A of the high-dose group decreased dramatically. When cells were treated for 24 h, the A of the low-dose group was significantly lower than that of the control group, while the A of the middle-dose and high-dose groups was much lower than control.② When cells were treated for 24 h, the proliferation of middle-dose group decreased, and that of high-dose group decreased markedly. Conclusions:Sodium selenate can inhibit the viability and proliferation of PC-3 cells, and these actions occur in a dose-dependant manner.