PRLs Promote Spreading, Adhesion, and Proliferation of Human SW480 and SW620 Cells

IntroductionMetastasis is the leading cause of the death ofcancer patients.It represents a complex and multi-stepprocess including the detachment of tumor cells from aprimary cancer,the invasion into the surrounding tis-sue,the entry into the circulatory system,the reinva-sion,and the proliferation at a distant secondarysite[1,2].A wide variety of stimuli are known to be as-sociated with the metastasis of tumor cells,which in-clude cytokines,hormones,growth factors,and extra-cellular matrix pr…     

Restoration of full-length APC protein in SW480 colon cancer cells induces exosome-mediated secretion of DKK-4

Mutations in the adenomatous polyposis coli (APC) tumor suppressor gene are common in both inherited and sporadic forms of colorectal cancer (CRC), and are associated with dysregulated Wnt signaling. Colon carcinoma SW480 cells restored with stable expression of wild-type APC (SW480APC cells) exhibit attenuated Wnt signaling, and reduced tumorigenicity, including increased cell adhesion. We performed a comparative proteomic analysis of exosomes isolated from SW480 and SW480APC cells to examine the effects of restored APC on exosome protein expression. A salient finding of our study was the unique expression of the Wnt antagonist Dickkopf-related protein 4 (DKK4) in SW480APC, but not parental SW480 cell-derived exosomes. Upregulation of DKK4 in SW480APC cells was confirmed by semiquantitative RT-PCR, immunoblotting, and immunogold electron microscopy. Analysis of the DKK4 gene promoter by methylation-specific PCR revealed reduced methylation in SW480APC cells, while RT-PCR demonstrated the downregulation of DNMT-3a, compared to the parental cell line. Our discovery of exosome-mediated secretion of DKK4 opens up the possibility that exosomal DKK4 may be a mechanism used by epithelial colon cells to regulate Wnt signaling which is lost during CRC progression.     

New Hybrids Based on Curcumin and Resveratrol: Synthesis,Cytotoxicity and Antiproliferative Activity against Colorectal Cancer Cells

We synthesized twelve hybrids based on curcumin and resveratrol, and their structures were elucidated by spectroscopic analysis. The chemopreventive potential of these compounds was evaluated against SW480 human colon adenocarcinoma cells, its metastatic derivative SW620, along with the non-malignant CHO-K1 cell line. Among the tested compounds, hybrids 3e and 3i (for SW480) and 3a, 3e and 3k (for SW620) displayed the best cytotoxic activity with IC₅₀ values ranging from 11.52 ± 2.78 to 29.33 ± 4.73 µM for both cell lines, with selectivity indices (SI) higher than 1, after 48 h of treatment. Selectivity indices were even higher than those reported for the reference drug, 5-fluorouracil (SI = 0.96), the starting compound resveratrol (SI = 0.45) and the equimolar mixture of curcumin plus resveratrol (SI = 0.77). The previous hybrids showed good antiproliferative activity.     

HnRNP K and PDI marked response to chemotherapy to human colorectal cancer cells

5‐Fluorouracil has been the chemotherapy agent of first‐choice for colorectal cancer for many years, but since there are no proven predictors of a patient's response to therapy, all patients receive similar treatment. Consequently, identification of biomarkers for therapeutic effect is crucial for the development of novel therapeutic strategies. Two human colorectal cancer cell lines of different metastatic potential (LoVo and SW480) were studied. IC50 of 5‐FU for both cell lines were measured by 3‐(4,5‐dimethy‐lthiazol‐2‐yl)‐2,5‐diphenyltetrazolium assay and validated by cell cycle analysis. Then the cell lines were treated with 5‐FU at IC50 concentration and protein was extracted for 2‐DE. Differential protein spots were examined by MALDI‐TOF/TOF MS. The expression levels of the different proteins were further confirmed by Western blot and immunofluorescence analyses. Eleven proteins were identified. Expression of heterogeneous nuclear ribonucleoprotein K (hnRNP K) in LoVo cells was higher than in SW480 cells, while protein disulfide isomerase (PDI) displayed the opposite trend. After treatment with 5‐FU, the expression of hnRNP K in LoVo decreased more significantly than in SW480, while PDI in SW480 increased more significantly than in LoVo cells. Conclusion: hnRNP K and PDI in the two cell lines have different expression characteristics. The sensitivity to 5‐FU is not consistent in tumor progression. It may assist in development of novel treatment strategies for colorectal cancer metastasis.     

Spectroscopic characterization of human and mouse primary cells,cell lines and malignant cells

Fourier transform infrared (FTIR) spectroscopy is currently being developed as a new optical approach to the diagnosis and characterization of cell or tissue pathology. The advantage of FTIR microspectroscopy over conventional FTIR spectroscopy in the diagnosis of malignancies is that it facilitates inspection of restricted regions of the cell culture or tissue. In this study, we set out to evaluate FTIR microspectroscopy as a diagnostic tool for identifying retrovirus-induced malignancies. Our study showed significant and consistent differences between cultures of different types of cells of both mouse and human origin, i.e. primary fibroblast cells (one to two passages in cell culture), fibroblast cell lines and malignant cells transformed by murine sarcoma virus. An impressive decrease in the levels of phosphate and other metabolites was seen in malignant cells compared with primary cells. The levels of these metabolites in the cell lines were significantly lower than in the primary cells but higher than in the malignant cells. In addition, the peak attributed to the PO2- symmetric stretching mode at 1082 cm(-1) in primary cells shifted significantly to 1085 cm(-1) for the cell line and to 1087 cm(-1) for the malignant cells. These differences taken together with differences in the shapes of various bands throughout the spectrum strongly support the possibility of developing FTIR microspectroscopy for the detection and study of malignant--and possibly premalignant--cells.     

Photodynamic Therapy with Zinc Phthalocyanine Inhibits the Stemness and Development of Colorectal Cancer: Time to Overcome the Challenging Barriers?

Photodynamic therapy (PDT) is a light-based cancer therapy approach that has shown promising results in treating various malignancies. Growing evidence indicates that cancer stem cells (CSCs) are implicated in tumor recurrence, metastasis, and cancer therapy resistance in colorectal cancer (CRC); thus, targeting these cells can ameliorate the prognosis of affected patients. Based on our bioinformatics results, SOX2 overexpression is significantly associated with inferior disease-specific survival and worsened the progression-free interval of CRC patients. Our results demonstrate that zinc phthalocyanine (ZnPc)-PDT with 12 J/cm² or 24 J/cm² irradiation can substantially decrease tumor migration via downregulating MMP9 and ROCK1 and inhibit the clonogenicity of SW480 cells via downregulating CD44 and SOX2. Despite inhibiting clonogenicity, ZnPc-PDT with 12 J/cm² irradiation fails to downregulate CD44 expression in SW480 cells. Our results indicate that ZnPc-PDT with 12 J/cm² or 24 J/cm² irradiation can substantially reduce the cell viability of SW480 cells and stimulate autophagy in the tumoral cells. Moreover, our results show that ZnPc-PDT with 12 J/cm² or 24 J/cm² irradiation can substantially arrest the cell cycle at the sub-G1 level, stimulate the intrinsic apoptosis pathway via upregulating caspase-3 and caspase-9 and downregulating Bcl-2. Indeed, our bioinformatics results show considerable interactions between the studied CSC-related genes with the studied migration- and apoptosis-related genes. Collectively, the current study highlights the potential role of ZnPc-PDT in inhibiting stemness and CRC development, which can ameliorate the prognosis of CRC patients.     

Oral cancer diagnostics based on infrared spectral markers and wax physisorption kinetics

Infrared microspectroscopy is an emerging approach for disease analysis owing to its capability for in situ chemical characterization of pathological processes. Synchrotron-based infrared microspectroscopy (SR-IMS) provides ultra-high spatial resolution for profiling biochemical events associated with disease progression. Spectral alterations were observed in cultured oral cells derived from healthy, precancerous, primary, and metastatic cancers. An innovative wax-physisorption-based kinetic FTIR imaging method for the detection of oral precancer and cancer was demonstrated successfully. The approach is based on determining the residual amount of paraffin wax (C₂₅H₅₂) or beeswax (C₄₆H₉₂O₂) on a sample surface after xylene washing. This amount is used as a signpost of the degree of physisorption that altered during malignant transformation. The results of linear discriminant analysis (LDA) of oral cell lines indicated that the methylene (CH₂) and methyl group (CH₃) stretching vibrations in the range of 3,000–2,800 cm^?1 have the highest accuracy rate (89.6 %) to discriminate the healthy keratinocytes (NHOK) from cancer cells. The results of wax-physisorption-based FTIR imaging showed a stronger physisorption with beeswax in oral precancerous and cancer cells as compared with that of NHOK, which showed a strong capability with paraffin wax. The infrared kinetic study of oral cavity tissue showed a consistency in the wax physisorption of the cell lines. On the basis of our findings, these results show the potential use of wax-physisorption-based kinetic FTIR imaging for the early screening of oral cancer lesions and the chemical changes during oral carcinogenesis.

Proapoptotic Effect and Molecular Docking Analysis of Curcumin–Resveratrol Hybrids in Colorectal Cancer Chemoprevention

Different hybrids based on curcumin and resveratrol were previously synthesized and characterized by spectroscopic techniques. The most active molecules (3a, 3e, 3i, and 3k) were evaluated in vitro as an approach to determine the possible mechanism of action of the hybrids. The results indicated that the evaluated curcumin/resveratrol hybrids induce mitochondrial instability in SW620 and SW480 cells. Moreover, these molecules caused a loss in membrane integrity, suggesting an apoptotic process mediated by caspases after the treatment with compounds 3i (SW480) and 3k (SW620). In addition, the results suggest that the mechanism of action of the hybrids could be independent of the p53 status. Furthermore, hybrids 3e and 3i caused G0/G1 phase arrest, which highlights the potential of these molecules not only as cytotoxic but also as cytostatic compounds. Hybrids 3e and 3i caused a negative modulation of the matrix metalloproteinase 7 (MMP7) on SW480 cells. These curcumin resveratrol hybrids could be potential candidates for further investigations in the search for potential chemopreventive agents, even in those cases with resistance to conventional chemotherapy because of the lack of p53 expression or function. Molecular docking simulations showed that compounds 3e, 3i, and 3k bind efficiently to proapoptotic human caspases 3/7 proteins, as well as human MMP-7 and p53, which, in turn, could explain at the molecular level the in vitro cytotoxic effect of these compounds in SW480 and SW620 colon cancer cell lines.     

First isolation of sesquiterpenes and flavonoids from Zingiber spectabile and identification of zerumbone as the major cell growth inhibitory component

Nine sesquiterpenes and eight flavonoids were isolated from Zingiber spectabile for the first time. Structures were determined by spectroscopic methods. The major compound zerumbone (1) was found to be the most active (IC(50) 13 microg mL(-1)) in cell growth inhibitory assay against colon carcinoma SW480 cells.     

A decade of vibrational micro-spectroscopy of human cells and tissue (1994-2004)

Instrumentation used in infrared microspectroscopy (IR-MSP) permits the acquisition of spectra from samples as small as 100 pg (10(-10) g), and as small as 1 pg for Raman microspectroscopy (RA-MSP). This, in turn, allows the acquisition of spectral data from objects as small as fractions of human cells, and of small regions of microtome tissue sections. Since vibrational spectroscopy is exquisitely sensitive to the biochemical composition of the sample, and variations therein, it is possible to monitor metabolic processes in tissue and cells, and to construct spectral maps based on thousands of IR spectra collected from pixels of tissue. These images, in turn, reveal information on tissue structure, distribution of cellular components, metabolic activity and state of health of cells and tissue.     

Modifying infrared scattering effects of single yeast cells with plasmonic metal mesh

The scattering effects in the infrared (IR) spectra of single, isolated bread yeast cells (Saccharomyces cerevisiae) on a ZnSe substrate and in metal microchannels have been probed by Fourier transform infrared imaging microspectroscopy. Absolute extinction [(3.4±0.6)×10(-7) cm(2) at 3178 cm(-1)], scattering, and absorption cross sections for a single yeast cell and a vibrational absorption spectrum have been determined by comparing it to the scattering properties of single, isolated, latex microspheres (polystyrene, 5.0 μm in diameter) on ZnSe, which are well modeled by the Mie scattering theory. Single yeast cells were then placed into the holes of the IR plasmonic mesh, i.e., metal films with arrays of subwavelength holes, yielding "scatter-free" IR absorption spectra, which have undistorted vibrational lineshapes and a rising generic IR absorption baseline. Absolute extinction, scattering, and absorption spectral profiles were determined for a single, ellipsoidal yeast cell to characterize the interplay of these effects.     

Spartium junceum aromatic water: chemical composition and antitumor activity

The purpose of this study was to analyse the chemical composition of Spartium junceum L. (also known as Spanish Broom) aromatic water and to evaluate its cytotoxic activity against a series of human cancer cell lines (melanoma: RPMI 7932; leukemia: K562; breast cancer cell: MCF7-Bart and MCF7-ICLC, colon adenocarcinoma: SW480). The results show that the aromatic water was cytotoxic toward the tumor cell lines analyzed (RPMI 7932, K562, MCF7-Bart, MCF7-ICLC, SW480), while it did not appreciably alter the viability of normal keratinocytes (NCTC 2544) suggesting its potential use as an antitumor agent for cancer treatment and/or prevention.     

Synthesis,characterization, and antitumor activity of novel tumor-targeted platinum(IV) complexes

Four tumor-targeted platinum(IV) complexes with ammonia and cyclohexylamine as the carrier groups and biotin as the axial group were designed, synthesized, and characterized. In vitro evaluation of the antitumor activity of complexes C1–C4 against lung cancer cells (A549), liver cancer cells (SMMC-7721), breast cancer cells (MCF-7), and colon cancer cells (SW480) was carried out. Complex C3 had the best cellular activity. Compared with cisplatin, complex C3 showed good anticancer activity against A549 cell line,complex C3 (6.34±0.44) is 3 times more cytotoxic than cisplatin (19.40±0.71),and against MCF-7 cell line complex C3 (4.22±0.11) is 5.4 times more cytotoxic than cisplatin (22.96±0.58), and against SW480 cell line complex C3 (6.65±0.60) is 3.4 times more cytotoxic than cisplatin (23.15±0.22). (Table 1) Axial chloride increased the redox power of complex C3 to increase the intercellular accumulation and the introduction of mixed amine had the ability to overcome cisplatin resistance. Complex C3 works best on MCF-7, then SW480, A549, and SMMC-7721. Thus, complex C3 is targeted by the axial introduction of biotin.     

Integration of intra- and extravasation in one cell-based microfluidic chip for the study of cancer metastasis

Most studies of cancer metastasis focus on cancer cell invasion utilizing adhesion assays that are performed independently, and are thus limited in their ability to mimic complex cancer metastasis on a chip. Here we report the development of an integrated cell-based microfluidic chip for intra- and extravasation that combines two assays on one chip for the study of the complex cascade of cancer metastasis. This device consists of two parts; one is an intravasation chamber for the three-dimensional (3-D) culture of cancer cells using a Matrigel matrix, and the other is an extravasation chamber for the detection of metastasized cancer cells by adhesion molecules expressed by epithelial cells. In this novel system, the intravasation and extravasation processes of cancer metastasis can be studied simultaneously using four screw valves. Metastatic LOVO and non-metastatic SW480 cells were used in this study, and the invasion of LOVOs was found to be higher compared to SW480. In contrast, invasion of cells treated with metalloproteinase (MMP) inhibitors decreased within the intravasation chamber. Degraded cancer cells from the intravasation chamber were detected within the extravasation chamber under physiological conditions of shear stress, and differences in binding efficiency were also detected when CA19-9 antibody, an inhibitor of cancer cell adhesion, was used to treat degraded cancer cells. Our results support the potential usefulness of this new 3D cell-based microfluidic system as a drug screening tool to select targets for the development of new drugs and to verify their effectiveness.     

基于亲和色谱的肺癌细胞磷酸化蛋白质组研究及其应用

磷酸化是蛋白质翻译后修饰的重要形式之一,其异常往往会导致细胞内信号通路的紊乱和疾病的发生。固定化金属离子亲和色谱(IMAC)是磷酸化肽段的高效富集技术,在磷酸化蛋白质组研究方面应用广泛。该研究以金属钛离子(Ti⁴⁺)螯合IMAC材料(Ti⁴⁺-IMAC)为载体,进行磷酸化肽段富集。比较了10 μm Ti⁴⁺-IMAC通过振荡法和固相萃取法(SPE)富集磷酸肽的效果,发现振荡法可以富集到更多的磷酸肽;对比了两种尺寸(10 μm和30 μm)Ti⁴⁺-IMAC在磷酸化肽段富集中的差异,发现小尺寸材料富集效果更佳。进一步采用优化的策略比较了不同转移能力肺癌细胞的磷酸化蛋白质组,免标记定量蛋白质组学结果表明,优化的Ti⁴⁺-IMAC方法可以从正常的肺成纤维细胞MRC5、低转移肺癌细胞95C和高转移肺癌细胞95D中分别鉴定到510、863和1108种磷酸化蛋白质,其中317种为3组所共有。该研究共鉴定到1268种磷酸化蛋白质上的7560个磷酸化位点,其中1130个为差异磷酸化位点,文献报道显示部分异常表达的激酶与癌症转移密切相关。通过生信对比分析发现,异常表达的磷酸化蛋白质主要与细胞侵袭、迁移和死亡等细胞迁移方面的功能有关。通过优化磷酸化肽富集策略,初步阐明了磷酸化蛋白质网络的异常与肺癌转移之间的相关性,该方法有望用于肺癌进展相关的磷酸化位点、磷酸化蛋白质及其信号通路研究。     

Identification of secondary metabolites in medicinal and spice plants by NIR-FT-Raman microspectroscopic mapping

This paper demonstrates the special potential of vibrational NIR FT Raman microspectroscopy for the study of fennel fruits, chamomile inflorescence and curcuma roots to obtain detailed information about their microstructure and chemical composition. Microscopic Raman maps of fennel fruits demonstrate that anethole, which is the main essential oil component, is present in the whole mericarp with highest concentration at the top of the fruit. In situ measurements obtained of the essential oil cells are dominated by two bands observed at 1657 cm(-1) and 1609 cm(-1) which are characteristic for anethole. Raman images of chamomile inflorescence show that spiroethers, identified by significant bands between 2150 and 2250 cm(-1), are accumulated in the middle part of the flower head. Due to the intense curcumin bands in the Raman spectrum of curcuma root, the distribution of this dyeing substance can be clearly determined; highest concentration of curcumin was observed on the core of the root.     

Synthesis and biological evaluation of novel acenaphthene derivatives as potential antitumor agents

Twelve novel acenaphthene derivatives have been synthesized. The structures of all compounds were confirmed by 1H-NMR, MS and elemental analysis. Their antitumor activities were evaluated in six human solid tumor cell lines, namely non-small cell lung cancer (H460), human colon adenocarcinoma (SW480), human breast cancer cell (MDA-MB-468 and SKRB-3), human melanoma cell (A375) and human pancreatic cancer (BxPC-3). Among them, compound 3c shows the best antitumor activity against SKRB-3 cell line, as high as the positive control adriamycin.     

Two new compounds from Melodinus suaveolens

Two new compounds, 19-hydroxy-melodinine K (1) and melodiside (2), and 25 known compounds were isolated from leaves and twigs of Melodinus suaveolens. Their structures were elucidated based on 1- and 2-D NMR, FTIR, UV and MS spectroscopic data. 19-hydroxy-melodinine K showed cytotoxic activity against MDA-MB-231 breast, BCG-823 gastric, SW480 colon and Hela cancer cells.     

Synthesis method-dependent photothermal effects of colloidal solutions of platinum nanoparticles used in photothermal anticancer therapy

One of the most common anticancer therapies is photothermal therapy (PTT). The effectiveness of PTT depends on the photosensitizer being a molecule which is toxic for the cancer cells after electromagnetic wave irradiation. Therefore, a simulation of PTT was performed in this work on two colon cancer cells (SW480 and SW620) using platinum nanoparticles (Pt NPs). Interestingly, in the literature the dependence between the synthesis method and the photothermal properties of Pt NPs was not discussed. Consequently, in this paper, we evaluated the photothermal properties of Pt NPs synthesized by two different methods: polyol (PtI NPs) and green chemistry (PtII NPs). Scanning transmission electron microscopy revealed that the size of both Pt NPs obtained was 2 nm, the NPs were not agglomerated, and that the PtII NPs were distributed on green tea supports. The selected area electron diffraction and X-ray diffraction analysis confirmed the crystallinity of both types of Pt NPs. Fourier-transform infrared (FTIR) spectrum of the PtII NPs showed interactions between the NPs and stretching modes for C=O groups from flavonoids and polyphenols. Therefore, these chemical compounds could be responsible for reducing Pt⁴⁺ ions to Pt⁰. Moreover, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay showed that the PtII NPs exhibited 10% and 20% better cytotoxicity effect on SW480 and SW620 cells, than PtI NPs. The viability of cancer cells decreased when Pt NPs were used in PTT. The highest percentage of dead cells (82%) was observed for PtII NPs and 650-nm laser irradiation. FTIR and Raman spectroscopy showed structural changes induced by both Pt NPs and laser irradiation of cells in the range corresponding to levels of DNA, phospholipids, proteins, and lipids. Moreover, the calculated photothermal conversion efficiency showed that the value of this parameter is around 35%, regardless of the synthesis method and used wavelengths.