海参提取物TBL-12抑制激素非依赖性前列腺癌细胞的增殖和转移
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摘要
目的探讨TBL-12在体外对激素非依赖性前列腺癌细胞增殖、迁移和侵袭功能的影响及其可能的机制。方法采用不同浓度(30、60、90μg/mL)的TBL-12处理PC-3和DU145前列腺癌细胞,分别用Cell Counting Kit-8法(CCK-8法)、划痕实验、Transwell侵袭实验检测TBL-12对前列腺癌细胞增殖、迁移、侵袭能力的影响,通过明胶酶谱检测基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)的酶活性。结果与对照组相比,TBL-12能显著抑制前列腺癌细胞增殖、迁移和侵袭能力,并呈剂量依赖性(P<0.05)。30μg/mL TBL-12能显著抑制前列腺癌细胞增殖和集落形成以及肿瘤转移(P<0.05)。90μg/mL的TBL-12处理后,PC-3细胞的集落形成率降至(8±3)%,迁移率由(53±11)%降为(12±4)%,侵袭细胞数由每个视野(1 085±101)个细胞减少到(29±18)个;而DU145细胞的集落形成率降至(5±2)%,细胞迁移率由(54±6)%降为(9±1)%,侵袭细胞数由每个视野(516±38)个减少到(10±9)个。明胶酶谱检测发现TBL-12可降低MMP-2、MMP-9的酶活性(P<0.05),90μg/mL的TBL-12处理后,PC-3细胞中MMP-9的酶活性降至(36±5)%,MMP-2降为(43±8)%;DU145细胞中MMP-9的酶活性降至(30±10)%,MMP-2降为(40±6)%。结论 TBL-12能显著抑制激素非依赖性前列腺癌细胞在体外的增殖、迁移和侵袭能力,具有潜在的治疗前列腺癌的临床应用价值。
Objective This study intends to explore the effects of TBL-12 on the proliferation,migration,invasion and apoptosis of prostate cancer PC3 and DU145 cells in vitro and its possible mechanism.MethodsThe prostate cancer cells were treated with different concentration of TBL-12(30,60,90μg/ml).Cell Counting Kit-8(CCK-8),scratch test and transwell invasion test were used to detect the effects of TBL-12 on the proliferation,migration,invasion of prostate cancer cells,respectively.Enzyme activities of matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9)were detected by Gelatin Zymography.ResultsCompared with the control group,TBL-12 could significantly inhibit the proliferation,migration and invasion ability of prostate cancer cells in a dose-dependent manner(P<0.05).30μg/mL TBL-12 also significantly inhibited colony formation and metastasis of prostate cancer cells(P<0.05).When treated with 90μg/mL TBL-12,the colonization rate of PC-3 cells decreased to(8±3)%,the migration rate decreased from(53±11)%to(12±4)%,the 262 number of invasive cells decreased from(1 085±101)cells per field to(29±18)cells,the colonization rate of DU145 cells decreased to(5±2)%,the cell migration rate decreased from(54±6)%to(9±1)%,the number of invasive cells decreased from(516±38)to(10±9)per field of vision.Gelatin Zymography showed that TBL-12 could decrease the activity of MMP-2and MMP-9(P<0.05).After 90μg/mL TBL-12 treatment,the activity of MMP-9 in PC-3 cells decreased to(36±5)%,MMP-2decreased to(43±8)%,MMP-9 decreased to(30±10)%and MMP-2 to(40±6)%in DU145 cells.Conclusion TBL-12 could significantly inhibit the proliferation,migration and invasion of hormone refractory prostate cancer cells in vitro;these results showed that TBL-12 had potential clinical application value in the treatment of prostate cancer.
Objective This study intends to explore the effects of TBL-12 on the proliferation,migration,invasion and apoptosis of prostate cancer PC3 and DU145 cells in vitro and its possible mechanism.MethodsThe prostate cancer cells were treated with different concentration of TBL-12(30,60,90μg/ml).Cell Counting Kit-8(CCK-8),scratch test and transwell invasion test were used to detect the effects of TBL-12 on the proliferation,migration,invasion of prostate cancer cells,respectively.Enzyme activities of matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9)were detected by Gelatin Zymography.ResultsCompared with the control group,TBL-12 could significantly inhibit the proliferation,migration and invasion ability of prostate cancer cells in a dose-dependent manner(P<0.05).30μg/mL TBL-12 also significantly inhibited colony formation and metastasis of prostate cancer cells(P<0.05).When treated with 90μg/mL TBL-12,the colonization rate of PC-3 cells decreased to(8±3)%,the migration rate decreased from(53±11)%to(12±4)%,the 262 number of invasive cells decreased from(1 085±101)cells per field to(29±18)cells,the colonization rate of DU145 cells decreased to(5±2)%,the cell migration rate decreased from(54±6)%to(9±1)%,the number of invasive cells decreased from(516±38)to(10±9)per field of vision.Gelatin Zymography showed that TBL-12 could decrease the activity of MMP-2and MMP-9(P<0.05).After 90μg/mL TBL-12 treatment,the activity of MMP-9 in PC-3 cells decreased to(36±5)%,MMP-2decreased to(43±8)%,MMP-9 decreased to(30±10)%and MMP-2 to(40±6)%in DU145 cells.Conclusion TBL-12 could significantly inhibit the proliferation,migration and invasion of hormone refractory prostate cancer cells in vitro;these results showed that TBL-12 had potential clinical application value in the treatment of prostate cancer.
引文
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[15]Lokeshwar BL,Selzer MG,Block NL,et al.Secretion of matrix metalloproteinases and their inhibitors(tissue inhibitor of metalloproteinases)by human prostate in explant cultures:reduced tissue inhibitor of metalloproteinase secretion by malignant tissues[J].Cancer Res,1993,53(19):4493-4498.
[16]Shen KH,Hung SH,Yin LT,et al.Acacetin,a flavonoid,inhibits the invasion and migration of human prostate cancer DU145 cells via inactivation of the p38 MAPK signaling pathway[J].Mol Cell Biochem,2010,333(1-2):279-291.
[17]Xu S,Zhou W,Ge J,et al.Prostaglandin E2 receptor EP4 is involved in the cell growth and invasion of prostate cancer via the cAMP-PKA/PI3K-Akt signaling pathway[J].Mol Med Rep,2018,17(3):4702-4712.
[18]Huang H,Du T,Xu G,et al.Matrine suppresses invasion of castration-resistant prostate cancer cells by downregulating MMP-2/9 via NF-κB signaling pathway[J].Int J Oncol,2016,50(2):640-648.
[2]Bubendorf L,Schopfer A,Wagner U,et al.Metastatic patterns of prostate cancer:an autopsy study of 1,589 patients[J].Hum Pathol,2000,31(5):578-583.
[3]张永勤,常海燕.海参内脏组成成分结构、功能及其应用研究进展[J].食品工业科技,2014,35(13):382-386.
[4]Chari A,Mazumder A,Lau K,et al.A phase II trial of TBL-12sea cucumber extract in patients with untreated asymptomatic myeloma[J].Br J Haematol,2018,180(2):296-298.
[5]贺晶,孙延霞,鲍慧.miR-145对宫颈癌细胞增殖、迁移和侵袭的影响[J].热带医学杂志,2018,18(11):1409-1413.
[6]周凯,徐霖,袁磊,等.海参提取物TBL-12对胃癌HGC-27细胞增殖、迁移、侵袭和凋亡的影响[J].热带医学杂志,2018,18(3):302-306.
[7]Zhang JJ,Zhu KQ.A novel antitumor compound nobiliside Disolated from sea cucumber(Holothuria nobilis Selenka)[J].Exp Ther Med,2017,14(2):1653-1658.
[8]Cuong NX,Vien LT,Hoang L,et al.Cytotoxic triterpene diglycosides from the sea cucumber Stichopus horrens[J].Bioorg Med Chem Lett,2017,27(13):2939-2942.
[9]毛晓鹏,陈俊星,莫承强,等.多西紫杉醇联合二烯丙三硫协同诱导前列腺癌细胞凋亡的研究[J].中山大学学报:医学科学版,2012,33(6):728-732.
[10]Mignatti P,Rifkin DB.Biology and biochemistry of proteinases in tumor invasion[J].Physiol Rev,1993,73(1):161-195.
[11]Nunn SE,Peehl DM,Cohen P.Acid-activated insulin-like growth factor binding protein protease activity of cathepsin D in normal and malignant prostatic epithelial cells and seminal plasma[J].J Cell Physiol,1997,171(2):196-204.
[12]Webber MM,Waghray A,Bello D,et al.Mini review:Proteases and invasion in human prostate epithelial cell lines:Implications in prostate cancer prevention and intervention[J].Radiation Oncology Investigations,1996,3(6):358-362.
[13]Stearns ME,Wang M,Res C,et al.Type IV collagenase(Mr72,000)expression in human[J].Cancer Res,1993,53(4):878-883.
[14]Wood M,Fudge K,Mohler JL,et al.In situ hybridization studies of metalloproteinases 2 and 9 and TIMP-1 and TIMP-2expression in human prostate cancer[J].Clin Exp Metastasis,1997,15(3):246-258.
[15]Lokeshwar BL,Selzer MG,Block NL,et al.Secretion of matrix metalloproteinases and their inhibitors(tissue inhibitor of metalloproteinases)by human prostate in explant cultures:reduced tissue inhibitor of metalloproteinase secretion by malignant tissues[J].Cancer Res,1993,53(19):4493-4498.
[16]Shen KH,Hung SH,Yin LT,et al.Acacetin,a flavonoid,inhibits the invasion and migration of human prostate cancer DU145 cells via inactivation of the p38 MAPK signaling pathway[J].Mol Cell Biochem,2010,333(1-2):279-291.
[17]Xu S,Zhou W,Ge J,et al.Prostaglandin E2 receptor EP4 is involved in the cell growth and invasion of prostate cancer via the cAMP-PKA/PI3K-Akt signaling pathway[J].Mol Med Rep,2018,17(3):4702-4712.
[18]Huang H,Du T,Xu G,et al.Matrine suppresses invasion of castration-resistant prostate cancer cells by downregulating MMP-2/9 via NF-κB signaling pathway[J].Int J Oncol,2016,50(2):640-648.