Evidence base and patients' perspective - World Journal of ...

More documents

Recommendations

Info

A 0��9 B Mean OD of colonosphere HFP C 0��8 0��7 0��6 0��5 0��4 0��3 0��2 0��1 0 DMSO Oxaliplatin Nigericin DMSO Oxaliplatin Nigericin Figure 3 Effects of nigericin and oxaliplatin on sphere- or colony-forming ability of cancer stem cells. A: Mean OD of sphere-forming HT29 cells after different treatments was assayed by methyl thiazolyl tetrazolium assay. Bars denote SD (n = 5); B: Numbers of colonies formed by HT29 cells after treatment in independent experiments. Bars denote SD (n = 5); C: Phase-contrast images of colonies formed in soft agar assays after treatment. DMSO: Dimethylsulfoxide; HPF: High-power field. form colonies under anchorage-independent conditions in a standard soft agar assay after 14 d in culture, relative to the control group (1.66 ± 0.57 vs 7 ± 1.15, P < 0.05), whereas the colony numbers were higher in oxaliplatin group relative to the vehicle-treated controls (14.33 ± 0.57 vs 7 ± 1.15, P < 0.05) (Figure 3B and C). Up-regulation of E-cadherin and downregulation of vimentin in CRC cells after nigericin treatment E-cadherin, encoded by the CDH1 gene, has dual functions in epithelial cells: as a cell-cell adhesion molecule and as a negative regulator of the canonical WNT signaling cascade; in particular, of its central mediator β-catenin. E-cadherin downregulation in mammalian cell systems is sufficient to trigger EMT [20] . Gupta et al [12] have reported that nigericin preferentially kills cells that have undergone EMT. In colorectal carcinomas, the embryonic EMT is activated during tumor invasion in disseminating cancer cells [21] . Characteristic of these cells is a loss of E-cadherin expression. We detected the expression of epithelial marker (Ecadherin) and mesenchymal marker (vimentin) of cells treated with nigericin and oxaliplatin to ascertain the effects of diverse compounds on EMT. As shown in Figure 4A and B, nigericin induced an increase in expression of E-cadherin and a decrease in expression of vimentin relative to vehicle controls. In contrast, the expression of E-cadherin in the cells treated with oxaliplatin was downregulated in contrast to vehicle WJG|www.wjgnet.com Number of colonies 16 14 12 10 8 6 4 2 0 controls; correspondingly, oxaliplatin treatment upregulated the expression level of vimentin. The data from real-time PCR showed similar results to immunocytochemistry and Western blotting (Figure 4C). DISCUSSION Zhou HM et al �� Nigericin and colorectal cancer DMSO Oxaliplatin Nigericin Significant progress has been made in understanding the molecular pathogenesis, diagnosis (hereditary and sporadic), and treatment of CRC. Despite the use of active targeted drugs for treatment of metastatic CRC in the past decade, and improvement of overall survival to nearly 2 years for nonresectable disease, the cure rate remains low [22] . 5-Fluorouracil and oxaliplatin formed the mainstay of chemotherapeutic regimens for metastatic CRC. Oxaliplatin covalently binds to DNA, forming platinum-DNA adducts that cause prolonged G2 arrest and inhibition of growth, which lead to apoptotic cell death [23] . There is a large body of evidence that tumor cells that are resistant to chemotherapy represent a subpopulation of cells from the primary tumor, which is molecularly and phenotypically distinct. These cells are referred to by several names, including tumor-initiating cells, tumorpromoting cells, or more commonly, CSCs [16] . EMT is a highly conserved cellular process during embryonic development and a pathogenic feature in tumorigenesis [10,24] . During the process of EMT, epithelial cells lose the expression of E-cadherin and other components of 2645 June 7, 2012|Volume 18|Issue 21|
A B Ecadherin Vimentin GAPDH Zhou HM et al �� Nigericin and colorectal cancer Ecadherin Vimentin epithelial cell junctions, adopt a mesenchymal cell phenotype, and acquire motility and invasive ability [25,26] . Furthermore, Mani et al [10] have induced EMT in nontumorigenic, immortalized human mammary epithelial cells (HMLEs) by ectopic expression of either the Twist or Snail transcription factors; these cells formed > 30-fold more mammospheres than did HMLEs infected with the corresponding control vector. They have concluded that the cells generated by EMT acquired yet another attribute of mammary stem cells. EMT, which enables cancer cell dissemination, also imparts a self-renewal capability to disseminating cancer cells. There is no consensus as to the exact criteria that define a CSC, because markers might vary according to the tumor type. In our study, we suggested CD133 as a marker of tumor-initiating cells of CRC [14-18] . We evaluated the effect of nigericin and oxaliplatin on CRC cell lines, including invasion and metastasis, and growth on colon cancer spheres, or colonospheres. From the results of cell viability and flow cytometry assays, we could see that nigericin specifically targets CD133 + cell WJG|www.wjgnet.com DMSO Oxaliplatin Nigericin DMSO Oxaliplatin Nigericin C Relative mRNA levels 3��5 3 2��5 2 1��5 1 0��5 0 E-cadherin Vimentin DMSO Oxaliplatin Nigericin Figure 4 Upregulation of E-cadherin and downregulation of vimentin in cancer stem cells after nigericin treatment. A: Immunofluorescence staining analysis of treated HT29 cells using epithelial E-cadherin and vimentin staining; B: E-cadherin and vimentin expression in HT29 cells after treatment, assayed by Western blotting; C: Real-time polymerase chain reaction analysis of E-cadherin and vimentin mRNA expression in HT29 cells after treatment with dimethylsulfoxide (DMSO) vehicle, oxaliplatin and nigericin. subpopulations within CRC cell lines. Moreover, nigericin induced inhibition of invasion and metastasis in HT29 cells. These effects may have been due to the fact that nigericin upregulated the expression of E-cadherin, while E-cadherin played an important role in cancer progression and EMT induction [27,28] . In a variety of human cancers, E-cadherin loss was closely related to poor prognosis, tumor progression, and metastasis [29,30] . Therefore, E-cadherin also could be a sign of drug efficiency of nigericin therapy in the future. Through analysis of the expression level of E-cadherin and vimentin, we may conclude that nigericin partly reverses EMT to affect the ability of CRC cells to invade and metastasize. We further evaluated the effects of nigericin treatment on the characteristics of CSC phenotype. The nigericin treatment group had a decreased number of spheres or colonies relative to the vehicle control group. Our data led us to hypothesize further that nigericin treatment suppresses EMT-generating cells with the properties of stem cells. This hypothesis needs further studies using animal 2646 June 7, 2012|Volume 18|Issue 21|
Page 1 and 2:
World Journal of Gastroenterology W
Page 3 and 4:
Phillip S Oates, Perth Ross C Smith
Page 5 and 6:
Deepak N Amarapurkar, Mumbai Abhiji
Page 7 and 8:
Trine Olsen, Tromsø Eyvind J Pauls
Page 9 and 10:
Conor P Delaney, Cleveland Sharon D
Page 11 and 12:
S Contents EDITORIAL FIELD OF VISIO
Page 13:
Contents ACKNOWLEDGMENTS APPENDIX A
Page 19: Cereda S et al . Adjuvant treatment
Page 22 and 23: DM, Sterling RK, Shiffman M, Topaz
Page 25 and 26: Selinger CP et al . Pregnancy relat
Page 31: Selinger CP et al . Pregnancy relat
Page 35 and 36: Ilan Y. Bacterial translocation in
Page 43: Ettreiki C et al . Iron prevents co
Page 46 and 47: A Macroscopir damage scores (AU) 7.
Page 48 and 49: A Total flora 12.5 B Log copy numbe
Page 51 and 52: Ettreiki C et al . Iron prevents co
Page 53: Online Submissions: http://www.wjgn
Page 57 and 58: A Fold changes over untreated cells
Page 59 and 60: A Migration Invasion B Migration In
Page 61 and 62: A miR-95 B Li WG et al �� Glarg
Page 64: Key words: Colorectal cancer; Niger
Page 67: A HT29 cells 100 C SW116 cells 40 P
Page 71 and 72: Zhou HM et al �� Nigericin and
Page 73 and 74: Terzin V et al . Serum IgG4 in auto
Page 75 and 76: Terzin V et al . Serum IgG4 in auto
Page 77 and 78: Online Submissions: http://www.wjgn
Page 79 and 80: Min YW et al . Sub-centimeter-sized
Page 85: Park CH et al . Family history and
Page 91: Online Submissions: http://www.wjgn
Page 96 and 97: genetic association study. BMC Canc
Page 98: ed ghrelin levels. World J Gastroen
Page 101 and 102: A Serum acylated ghrelin (pg/mL) B
Page 103 and 104: Yang YJ et al . H. pylori eradicati
Page 105 and 106: Online Submissions: http://www.wjgn
Page 107 and 108: Chen JG et al . Colorectal cancer s
Page 109 and 110: Chen JG et al . Colorectal cancer s
Page 111: Chen JG et al . Colorectal cancer s
Page 117 and 118: Qiao CX et al . Quality of life wit
Page 120:
primary liver cancer and one of the
Page 123 and 124:
A A 490 Li YH et al . GABRQ in HCC
Page 125 and 126:
Li YH et al . GABRQ in HCC prostate
Page 127 and 128:
Online Submissions: http://www.wjgn
Page 129 and 130:
Table 1 The main characteristics of
Page 131 and 132:
Shen Y et al . PTEN and cetuximab e
Page 133 and 134:
Shen Y et al . PTEN and cetuximab e
Page 135 and 136:
Yang XX et al . Aberrant methylatio
Page 137 and 138:
Relative expression 10 8 6 4 2 0 1.
Page 139 and 140:
E Methylation (%) F Methylation (%)
Page 141 and 142:
Yang XX et al . Aberrant methylatio
Page 143 and 144:
Di Nardo G et al . Octreotide in pe
Page 145 and 146:
Page 147 and 148:
A B Aniwan S et al . Infliximab for
Page 149 and 150:
Aniwan S et al . Infliximab for CD
Page 151 and 152:
Nishizawa T et al . Dual therapy fo
Page 153 and 154:
Nishizawa T et al . Dual therapy fo
Page 155 and 156:
Page 157 and 158:
Instructions to authors ISSN and EI
Page 159 and 160:
Instructions to authors AIM (no mor
Page 161 and 162:
Instructions to authors Guidelines
show all

Evidence base and patients' perspective - World Journal of ...

Create successful ePaper yourself

Delete template?

Save as template?