Epithelial-mesenchymal transition in residual gastric cells after chemotherapy: An in vitro study
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摘要:
目的 观察体外化疗能否诱导胃癌细胞发生上皮间质转化（epithelial mesenchymal transition，EMT）。 方法 使用5-Fu（30 μg/mL）对胃癌细胞株SGC7901进行4个疗程的体外化疗，残余细胞继续培养，得到能稳定传代的细胞SGC7901/Fu。比较SGC7901及SGC7901/Fu在细胞形态、EMT标记物、化疗耐药性、侵袭能力、肿瘤干细胞特性等方面的差异。 结果 与SGC7901比较，SGC7901/Fu呈间质细胞形态、上皮表型标记物表达下调、间质表型标记物表达上调。在SGC7901细胞及SGC7901/Fu细胞中，5-Fu的中效浓度（IC50）分别为（43.8±7.2）μg/mL及（64.6±5.5）μg/mL，穿过Transwell小室基底膜的细胞数分别为（51.4±8.7）个及（93.2±9.5）个，克隆形成率分别为5.2%±1.0%及13.2%±2.2%，CD44+/CD24-细胞亚群所占比例分别为4.13%±0.81%及7.97%± 0.50%，两者间的差异均具有统计学意义（P<0.05）。 结论 体外化疗后残余的胃癌细胞发生EMT，同时细胞侵袭能力增强、化疗耐药性升高，并获得了肿瘤干细胞特性。 Abstract:Objective To investigate if in vitro chemotherapy can induce the EMT progress in gastric cancer (GC) cells. Methods The GC cell line, SGC7901, was treated using 5-Fu at a concentration of 30 μg/mL. The residual cells after four cycles of 5-Fu therapy were named as SGC7901/Fu. The morphological changes and malignant biological features, including the invasiveness and clone formation ability and the characteristics of cancer stem cell and biomarkers of EMT between SGC7901 and SGC7901/Fu, were compared. Results The SGC7901/Fu cells displayed a mesenchymal appearance, decreased the expression of epithelial markers, and increased the expression of mesenchymal markers. The 50% inhibitory concentrations in the SGC7901/Fu and SGC7901 cells were (43.8±7.2) and (64.6±5.5) μg/mL, respectively. The number of cells that migrated through the basement-membrane of the Transwell chamber was 51.4±8.7 and 93.2±9.5, respectively. The rate of clone formation was 5.2%±1.0% and 13.2%±2.2%, respectively. The portions of the CD44+/CD24- cells were 4.13%±0.81% and 7.97%±0.50%, respectively. All differences were statistically significant (P< 0.05). Conclusion The residual GC cells underwent EMT progress after 5-Fu treatment, with increased chemoresistance and ability of invasiveness and acquired the property of cancer stem cells. -
Key words:
- gastric cancer /
- chemoresistance /
- epithelial-mesenchymal transition /
- cancer stem cell /
- 5-Fu
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图 1 SGC7901及SGC7901/Fu细胞形态比较（×200）
Figure 1. Morphological comparison between SGC7901 and SGC7901/Fu cells
A:SGC7901 cells;B:SGC7901/Fu cells
图 2 EMT及肿瘤干细胞相关基因的表达
Figure 2. Expression of EMT and cancer stem cell-related genes
A:Real-time reverse transcription-polymerase chain reaction test;B:Western blot test
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[1] Yang AD, Fan F, Camp ER, et al. Chronic oxaliplatin resistance in duces epithelial-to-mesenchymal transition in colorectal cancer cell lines[J]. Clin Cancer Res, 2006, 12(14 Pt 1): 4147-4153. [2] 徐志远, 戴朝六, 王宁. 塞来昔布对人肝癌细胞株增殖及凋亡的影响[J]. 肿瘤学杂志, 2008, 14(11): 902-905. https://www.cnki.com.cn/Article/CJFDTOTAL-XHON200811012.htm [3] Xu ZY, Ding SM, Zhou L, et al. FOXC1 contributes to microvascular invasion in primary hepatocellular carcinoma via regulating epitheli al-mesenchymal transition[J]. Int J Biol Sci, 2012, 8(8): 1130-1141. doi: 10.7150/ijbs.4769 [4] Taube JH, Herschkowitz JI, Komurov K, et al. Coreepitheli al-to-mesenchymal transition interactome gene-expression signature is associated with claudin-low and metaplastic breast cancer subtypes [J]. Proc Natl Acad Sci U S A, 2010, 107(35): 15449-15454. doi: 10.1073/pnas.1004900107 [5] Yang J, Weinberg RA. Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis[J]. Dev Cell, 2008, 14 (6): 818- 829. doi: 10.1016/j.devcel.2008.05.009 [6] Latifi A, Abubaker K, Castrechini N, et al. Cisplatin treatment of primary and metastatic epithelial ovarian carcinomas generates re sidual cells with mesenchymal stem cell-like profile[J]. J Cell Bio chem, 2011, 112(10): 2850-2864. [7] Basu D, Nguyen TT, Montone KT, et al. Evidence for mesenchy mal-like sub-populations within squamous cell carcinomas possess ing chemoresistance and phenotypic plasticity[J]. Oncogene, 2010, 29(29): 4170-4182. doi: 10.1038/onc.2010.170 [8] Maseki S, Ijichi K, Tanaka H, et al. Acquisition of EMT pheno type in the gefitinib-resistant cells of a head and neck squamous cell carcinoma cell line through Akt/GSK-3 β/snail signaling path way[J]. Br J Cancer, 2012, 106(6): 1196-1204. doi: 10.1038/bjc.2012.24 [9] Uchibori K, Kasamatsu A, Sunaga M, et al. Establishment and char acterization of two 5-fluorouracil-resistant hepatocellular carcino ma cell lines[J]. Int J Oncol, 2012, 40(4): 1005-1010. doi: 10.3892/ijo.2011.1300 [10] Zhang W, Feng M, Zheng G, et al. Chemoresistance to 5-fluoroura cil induces epithelial-mesenchymal transition via up-regulation of Snail in MCF-7 human breast cancer cells[J]. Biochem Biophys Res Commun, 2012, 417(2): 679-685. doi: 10.1016/j.bbrc.2011.11.142 [11] Marchini S, Fruscio R, Clivio L, et al. Resistance to platinum-based chemotherapy is associated with epithelial to mesenchymal transition in epithelial ovarian cancer[J]. Eur J Cancer, 2013, 49(2): 520-530. doi: 10.1016/j.ejca.2012.06.026 [12] Scheel C, Weinberg RA. Cancer stem cells and epithelial-mesenchy mal transition: concepts and molecular links[J]. Semin Cancer Biol, 2012, 22(5-6): 396-403. doi: 10.1016/j.semcancer.2012.04.001 [13] Hegde GV, de la Cruz C, Eastham-Anderson J, et al. Residual tu mor cells that drive disease relapse after chemotherapy do not have enhanced tumor initiating capacity[J]. PLoS One, 2012, 7(10): e45647. doi: 10.1371/journal.pone.0045647 -
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