瞬时受体电位香草酸亚型1在结肠癌组织中的表达及其对结肠癌细胞生物学行为的影响
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摘要
目的验证瞬时受体电位香草酸亚型1(TRPV1)在结肠癌组织及正常组织中的表达差异,并探讨其对结肠癌细胞生物学行为的影响。方法 TCGA数据库中发现结肠癌组织和正常组织中TRPV1蛋白存在差异表达,遂收集四川省人民医胃肠外科2018年5～8月的结肠癌组织、癌旁组织、正常组织共26例,使用免疫组织化学技术检测各标本中的TRPV1蛋白的表达情况;进一步在人结肠癌HCT116细胞系上进行功能实验,结肠癌HCT116细胞加入TRPV1激活剂培养后,使用CCK-8检测细胞活力验证增殖情况,Annexin V-FITC/PI双染检测细胞凋亡。结果在TCGA结肠癌细胞样本275例、正常组织样本349例中,结肠癌组织中TRPV1蛋白的表达明显低于其在正常组织中的表达。在我院收集的临床样本中进行验证,结果发现结肠癌组织中TRPV1蛋白的表达明显低于癌旁组织与正常组织,TRPV1蛋白在癌旁组织的表达介于癌组织和正常组织之间,表达差异有统计学意义。加入TRPV1激动剂培养结肠癌HCT116细胞后,CCK-8检测发现细胞增殖能力显著减弱,Annexin V-FITC/PI双染检测发现细胞凋亡增多。结论 TRPV1在结肠癌组织中表达降低,其生物学行为可减低结肠癌细胞增殖能力,促进结肠癌细胞凋亡。TRPV1在结肠癌细胞中存在表达抑制,具有一定的临床诊断意义。
Objective To verify the differences in the expression of transient receptor potential vanilloid 1( TRPV1) between colon cancer tissues and normal tissues,and to explore its effect on the biological behavior of colon cancer cells.Methods Different expression of TRPV1 protein in colon cancer tissues and normal tissues was found in the TCGA database.We collected 26 cases of colon cancer tissues,paracancerous tissues and normal tissues between May and August 2018 from Department of Gastrointestinal Surgery,Sichuan Provincial People's Hospital.Then,we detected TRPV1 protein expression in each specimen by using immunohistochemistry.After that,we further conducted functional experiments on human colon cancer HCT116 cell line. When the HCT116 cells were co-cultured with TRPV1 activator,we used CCK-8 to detect cell viability and proliferation and performed annexin V-FITC/PI double staining to detect apoptosis.Results In 275 cases of TCGA colon cancer samples and 349 cases of normal tissue samples,the expression of TRPV1 protein in colon cancer tissues was significantly lower than that in normal tissues.In our clinical samples,it was also found that the expression of TRPV1 protein in colon cancer tissues was significantly lower than that in paracancerous tissues and normal tissues while the expression of TRPV1 protein in paracancerous tissues was between cancer and normal tissues.The differences in TRPV1 expression were statistically significant. After adding TRPV1 agonist to culture colon cancer HCT116 cells,cell proliferation ability was significantly weakened and cell apoptosis was increased.Conclusion TRPV1 expression is reduced in colon cancer tissues. Its biological behavior can reduce the proliferation of colon cancer cells and promote the apoptosis of colon cancer cells.Therefore,TRPV1 has certain clinical diagnostic significance.
Objective To verify the differences in the expression of transient receptor potential vanilloid 1( TRPV1) between colon cancer tissues and normal tissues,and to explore its effect on the biological behavior of colon cancer cells.Methods Different expression of TRPV1 protein in colon cancer tissues and normal tissues was found in the TCGA database.We collected 26 cases of colon cancer tissues,paracancerous tissues and normal tissues between May and August 2018 from Department of Gastrointestinal Surgery,Sichuan Provincial People's Hospital.Then,we detected TRPV1 protein expression in each specimen by using immunohistochemistry.After that,we further conducted functional experiments on human colon cancer HCT116 cell line. When the HCT116 cells were co-cultured with TRPV1 activator,we used CCK-8 to detect cell viability and proliferation and performed annexin V-FITC/PI double staining to detect apoptosis.Results In 275 cases of TCGA colon cancer samples and 349 cases of normal tissue samples,the expression of TRPV1 protein in colon cancer tissues was significantly lower than that in normal tissues.In our clinical samples,it was also found that the expression of TRPV1 protein in colon cancer tissues was significantly lower than that in paracancerous tissues and normal tissues while the expression of TRPV1 protein in paracancerous tissues was between cancer and normal tissues.The differences in TRPV1 expression were statistically significant. After adding TRPV1 agonist to culture colon cancer HCT116 cells,cell proliferation ability was significantly weakened and cell apoptosis was increased.Conclusion TRPV1 expression is reduced in colon cancer tissues. Its biological behavior can reduce the proliferation of colon cancer cells and promote the apoptosis of colon cancer cells.Therefore,TRPV1 has certain clinical diagnostic significance.
引文
[1]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
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[3]Caterina MJ,Schumacher MA,Tominaga M,et al.The capsaicin receptor:a heat-activated ion channel in the pain pathway[J].Nature,1997,389(6653):816-824.
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[6]Marrone MC,Morabito A,Giustizieri M,et al. TRPV1 channels are critical brain inflammation detectors and neuropathic pain biomarkers in mice[J].Nat Commun,2017,8:15292.
[7]Prevarskaya N,Zhang L,Barritt G.TRP channels in cancer[J].Biochim Biophys Acta,2007,1772(8):937-946.
[8] Riera CE,Huising MO,Follett P,et al. TRPV1 pain receptors regulate longevity and metabolism by neuropeptide signaling[J].Cell,2014,157(5):1023-1036.
[9]石宏,何伟,王晓宇,等.TRPV1和ASIC3基因敲除小鼠的寿命观察[J].中国比较医学杂志,2014,24(10):78-81,89.
[10]Weber LV,Al-Refae K,Wlk G,et al.Expression and functionality of TRPV1 in breast cancer cells[J]. Breast Cancer(Dove Med Press),2016,8:243-252.
[11]Vercelli C,Barbero R,Cuniberti B,et al.Transient receptor potential vanilloid 1 expression and functionality in mcf-7 cells:a preliminary investigation[J].J Breast Cancer,2014,17(4):332-338.
[12]Vriens J,Janssens A,Prenen J,et al.TRPV channels and modulation by hepatocyte growth factor/scatter factor in human hepatoblastoma(Hep G2)cells[J].Cell Calcium,2004,36(1):19-28.
[13]Marincsák R,Tóth BI,Czifra G,et al. Increased expression of TRPV1 in squamous cell carcinoma of the human tongue[J]. Oral Dis,2009,15(5):328-335.
[14]Czifra G,Varga A,Nyeste K,et al.Increased expressions of cannabinoid receptor-1 and transient receptor potential vanilloid-1 in human prostate carcinoma[J]. J Cancer Res Clin Oncol,2009,135(4):507-514.
[15]Morelli MB,Amantini C,Nabissi M,et al. Cross-talk between alpha1D-adrenoceptors and transient receptor potential vanilloid type1 triggers prostate cancer cell proliferation[J]. BMC Cancer,2014,14(1):921.
[16]Hartel M,di MFF,Selvaggi F,et al.Vanilloids in pancreatic cancer:potential for chemotherapy and pain management[J]. Gut,2006,55(4):519-528.
[17]Vinuesa AG,Sancho R,García-Limones C,et al.Vanilloid receptor-1regulates neurogenic inflammation in colon and protects mice from colon cancer[J].Cancer Res,2012,72(7):1705-1716.
[18]de Jong PR,Takahashi N,Harris AR,et al.Ion channel TRPV1-dependent activation of PTP1B suppresses EGFR-associated intestinal tumorigenesis[J].J Clin Invest,2014,124(9):3793-806.
[19]Fernandes ES,Fernandes MA,Keeble JE. The functions of TRPA1and TRPV1:moving away from sensory nerves[J].Br J Pharmacol,2012,166(2):510-521.
[20]BarthóL,BenkóR,Patacchini R,et al. Effects of capsaicin on visceral smooth muscle:a valuable tool for sensory neurotransmitter identification[J].Eur J Pharmacol,2004,500(1-3):143-157.
[21]Holzer P.Efferent-like roles of afferent neurons in the gut:Blood flow regulation and tissue protection[J]. Auton Neurosci,2006,125(1-2):70-75.
[22]Holzer P.TRPV1 and the gut:from a tasty receptor for a painful vanilloid to a key player in hyperalgesia[J]. Eur J Pharmacol,2004,500(1-3):231-241.
[23]Waning J,Vriens J,Owsianik G,et al.A novel function of capsaicinsensitive TRPV1 channels:involvement in cell migration[J]. Cell Calcium,2007,42(1):17-25.
[24]Ganea D,Delgado M.Vasoactive intestinal peptide(VIP)and pituitary adenylate cyclase-activating polypeptide(PACAP)as modulators of both innate and adaptive immunity[J]. Crit Rev Oral Biol Med,2002,13(3):229-237.
[2]Siegel RL,Miller KD,Fedewa SA,et al.Colorectal cancer statistics,2017[J].CA Cancer J Clin,2017,67(3):177-193.
[3]Caterina MJ,Schumacher MA,Tominaga M,et al.The capsaicin receptor:a heat-activated ion channel in the pain pathway[J].Nature,1997,389(6653):816-824.
[4]Gunthorpe MJ,Benham CD,Randall A,et al.The diversity in the vanilloid(TRPV)receptor family of ion channels[J]. Trends Pharmacol Sci,2002,23(4):183-191.
[5]Ramsey IS,Delling M,Clapham DE.An introduction to TRP channels[J].Annu Rev Physiol,2006,68(68):619-647.
[6]Marrone MC,Morabito A,Giustizieri M,et al. TRPV1 channels are critical brain inflammation detectors and neuropathic pain biomarkers in mice[J].Nat Commun,2017,8:15292.
[7]Prevarskaya N,Zhang L,Barritt G.TRP channels in cancer[J].Biochim Biophys Acta,2007,1772(8):937-946.
[8] Riera CE,Huising MO,Follett P,et al. TRPV1 pain receptors regulate longevity and metabolism by neuropeptide signaling[J].Cell,2014,157(5):1023-1036.
[9]石宏,何伟,王晓宇,等.TRPV1和ASIC3基因敲除小鼠的寿命观察[J].中国比较医学杂志,2014,24(10):78-81,89.
[10]Weber LV,Al-Refae K,Wlk G,et al.Expression and functionality of TRPV1 in breast cancer cells[J]. Breast Cancer(Dove Med Press),2016,8:243-252.
[11]Vercelli C,Barbero R,Cuniberti B,et al.Transient receptor potential vanilloid 1 expression and functionality in mcf-7 cells:a preliminary investigation[J].J Breast Cancer,2014,17(4):332-338.
[12]Vriens J,Janssens A,Prenen J,et al.TRPV channels and modulation by hepatocyte growth factor/scatter factor in human hepatoblastoma(Hep G2)cells[J].Cell Calcium,2004,36(1):19-28.
[13]Marincsák R,Tóth BI,Czifra G,et al. Increased expression of TRPV1 in squamous cell carcinoma of the human tongue[J]. Oral Dis,2009,15(5):328-335.
[14]Czifra G,Varga A,Nyeste K,et al.Increased expressions of cannabinoid receptor-1 and transient receptor potential vanilloid-1 in human prostate carcinoma[J]. J Cancer Res Clin Oncol,2009,135(4):507-514.
[15]Morelli MB,Amantini C,Nabissi M,et al. Cross-talk between alpha1D-adrenoceptors and transient receptor potential vanilloid type1 triggers prostate cancer cell proliferation[J]. BMC Cancer,2014,14(1):921.
[16]Hartel M,di MFF,Selvaggi F,et al.Vanilloids in pancreatic cancer:potential for chemotherapy and pain management[J]. Gut,2006,55(4):519-528.
[17]Vinuesa AG,Sancho R,García-Limones C,et al.Vanilloid receptor-1regulates neurogenic inflammation in colon and protects mice from colon cancer[J].Cancer Res,2012,72(7):1705-1716.
[18]de Jong PR,Takahashi N,Harris AR,et al.Ion channel TRPV1-dependent activation of PTP1B suppresses EGFR-associated intestinal tumorigenesis[J].J Clin Invest,2014,124(9):3793-806.
[19]Fernandes ES,Fernandes MA,Keeble JE. The functions of TRPA1and TRPV1:moving away from sensory nerves[J].Br J Pharmacol,2012,166(2):510-521.
[20]BarthóL,BenkóR,Patacchini R,et al. Effects of capsaicin on visceral smooth muscle:a valuable tool for sensory neurotransmitter identification[J].Eur J Pharmacol,2004,500(1-3):143-157.
[21]Holzer P.Efferent-like roles of afferent neurons in the gut:Blood flow regulation and tissue protection[J]. Auton Neurosci,2006,125(1-2):70-75.
[22]Holzer P.TRPV1 and the gut:from a tasty receptor for a painful vanilloid to a key player in hyperalgesia[J]. Eur J Pharmacol,2004,500(1-3):231-241.
[23]Waning J,Vriens J,Owsianik G,et al.A novel function of capsaicinsensitive TRPV1 channels:involvement in cell migration[J]. Cell Calcium,2007,42(1):17-25.
[24]Ganea D,Delgado M.Vasoactive intestinal peptide(VIP)and pituitary adenylate cyclase-activating polypeptide(PACAP)as modulators of both innate and adaptive immunity[J]. Crit Rev Oral Biol Med,2002,13(3):229-237.