Reciprocal Interaction between Carcinoma-Associated Fibroblasts and Squamous Carcinoma Cells through Interleukin-1α Induces Cancer Progression

Crosstalk between cancer cells and carcinoma-associated fibroblasts (CAFs) has earned recognition as an interaction that plays a pivotal role in carcinogenesis. Thus, we attempted to clarify whether increase in the level of CAFs promotes cancer progression by proportionally enhancing the interaction between cancer cells and CAFs. We first analyzed clinical correlation between the levels of fibroblasts and cancer progression and found that the level of CAFs made a noticeable difference on the prognosis of patients with oral squamous cell carcinoma (OSCC). In vivo animal study also demonstrated that tumor volume depended on the dose of CAFs that was co-injected with OSCC cells. The same tendency was observed in an in vitro study. We also found that interleukin-1α (IL-1α) secreted from OSCC cells had dual effects on CAFs: IL-1α not only promoted the proliferation of CAFs but also upregulated the secretion of cytokines in CAFs such as CCL7, CXCL1, and IL-8. The induction activity of cytokine secretion by IL-1α surpassed that of proliferation in OSCC cells. In summary, we unraveled an important interactive mechanism of carcinogenesis: IL-1α released from carcinoma stimulates the proliferation of CAFs and the simultaneous increase in cytokine secretion from CAFs promotes cancer progression in human OSCC. On the basis of these findings, we propose that the level of CAFs is eligible for being selected as a prognostic factor that will be useful in routine diagnosis. We also propose that blockage of reciprocal interaction between cancer cells and CAFs will provide an insight for developing novel chemotherapeutic strategy.     

Heat shock factor 1 in cancer‐associated fibroblasts is a potential prognostic factor and drives progression of oral squamous cell carcinoma

Heat shock factor 1 (HSF1) is highly expressed in various malignancies and is a potential modulator of tumor progression. Emerging evidence suggests that HSF1 activation in stromal cells is closely related to poor patient prognosis. However, the role of HSF1 in oral squamous cell carcinoma (OSCC) remains elusive. We aimed to investigate the function of HSF1 in cancer‐associated fibroblasts (CAFs) of the tumor microenvironment (TME) and in tumor development. In the present study, we found that HSF1 was highly expressed in both CAFs and tumor cells, and was significantly correlated with poor prognosis and overall survival. Moreover, HSF1 overexpression in CAFs resulted in a fibroblast‐like phenotype of Cal27 cells, induced epithelial‐mesenchymal transition (EMT), and promoted proliferation, migration and invasion in Cal27 cells. HSF1 knockdown attenuated features of CAFs and reduced EMT, proliferation, migration and invasion in Cal27 cells. Furthermore, HSF1 in CAFs promoted tumor growth in nude mice. Taken together, these data suggest that HSF1 expression in CAFs drive OSCC progression, and could serve as an independent prognostic marker of patients with OSCC. Thus, HSF1 is a potent mediator of OSCC malignancy.     

Stromal fibroblasts influence oral squamous-cell carcinoma cell interactions with tenascin-C

In this study we identified tenascin-C (TN-C) and one of its integrin receptors, αvβ6, in oral squamous-cell carcinoma (SCC) specimens. Neither TN-C nor αvβ6 are expressed in normal oral mucosa. We also studied 2 human oral squamous-cell carcinoma cell lines: the highly invasive HSC-3 cells, and the poorly invasive SCC-25 cells. We determined that adhesion of these cells to TN-C involves both α2 and αv integrins. Migration on TN-C by oral SCC cells required fibroblast-conditioned medium and did not occur in its absence. This migration was blocked by anti-α2 and anti-αv antibodies and was partially inhibited by antibodies to hepatocyte growth factor, epidermal growth factor and transforming growth factor-β1. When seeded on TN-C, the poorly invasive SCC-25 cells formed αvβ6-positive focal contacts; the HSC-3 cells did not. HSC-3, SCC-25 and PTF cells secrete TN-C into the culture medium, as determined by Western blot. However, when HSC-3 cells were inoculated into the floor of the mouth of nude mice, only murine TN-C could be identified in the reactive stroma adjacent to the resulting tumor nests, demonstrating that in vivo, HSC-3 cells do not secrete TN-C. Our results demonstrate that αvβ6 and tenascin-C are neo-expressed in oral squamous-cell carcinoma, and that the tumor stromal environment is influential in oral SCC behavior. Int. J. Cancer 72:369–376, 1997. © 1997 Wiley-Liss, Inc.     

lncRNA MALAT1靶向miR-150对口腔鳞癌细胞增殖和凋亡的影响及分子机制

目的:研究MALAT1对口腔鳞癌细胞SCC-25增殖、凋亡的影响,并探讨其机制。方法:运用qRT-PCR检测人口腔鳞癌细胞SCC-25、人永生化口腔上皮细胞HIOEC中MALAT1和miR-150的mRNA表达;将si-con组（转染si-con）、si-MALAT1组（转染si-MALAT1）、miR-150组（转染miR-150 mimics）、miR-con组（转染miR-con）、si-MALAT1+anti-miR-con组（si-MALAT1和anti-miR-con共转染）、si-MALAT1+anti-miR-150组（si-MALAT1和anti-miR-150共转染）,均以脂质体法转染至SCC-25细胞;MTT法检测各组细胞的增殖;流式细胞术检测各组细胞的凋亡;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果:与人永生化口腔上皮HIOEC细胞（Normal组）相比,口腔鳞癌SCC-25细胞（Tumor组）中MALAT1表达显著上调,miR-150显著下调（P＜0.05）;敲减MALAT1、过表达miR-150均可抑制SCC-25细胞增殖,促进凋亡;MALAT1靶向miR-150。抑制miR-150逆转了敲减MALAT1对口腔鳞癌细胞的增殖抑制和凋亡促进作用。结论:MALAT1可促进口腔鳞癌细胞增殖并抑制凋亡,其机制可能与靶向miR-150有关,可为口腔鳞癌的治疗提供新靶点。     

Carcinoma-associated fibroblasts promotes the proliferation of a lingual carcinoma cell line by secreting keratinocyte growth factor

Carcinoma-associated fibroblasts (CAFs) have been confirmed to play an important role in the occurrence and development of many kinds of tumors. Regarding proliferation as one manifestation of malignance, the objective was to observe the effects of oral CAFs on the proliferation of oral squamous carcinoma cells (OSCC) and to explore the role of keratinocyte growth factor (KGF) in this process. The results showed that oral CAFs secreted a higher level of KGF than oral normal fibroblasts (NFs), and the conditioned medium of CAFs could increase the viability of carcinoma cells and promote more of them into G2 and S phase. However, after blocking with KGF antibody, the viability and cell cycle of Tca8113 cultured with CAFs conditioned medium changed to be similar with NFs control groups. It was concluded that CAFs could promote the proliferation of OSCC through secreting high levels of KGF. These findings support the use of carcinoma-associated fibroblasts as a novel target in anticancer therapy.     

Stromal–epithelial interactions modulate cross-talk between prolactin receptor and HER2/Neu in breast cancer

Prolactin (PRL) promotes the proliferation and survival of breast cancer cells in part via the transactivation of human epidermal growth factor receptor 2 (HER2), also known as Neu in rodents. A PRL receptor (PRLR) antagonist, G129R, has been developed, which indirectly inhibits the tyrosine phosphorylation of HER2 (p-HER2) in human breast cancer cell lines. In this study, we investigate the effects of cancer-associated fibroblasts (CAFs) upon this molecular cross-talk using tumor cells and CAFs derived from spontaneous mammary tumors of female MMTV-neu transgenic mice. Tumors were resected and cultured as small tumor chunks (~3 mm3) or were cultured in monolayer. G129R reduced tyrosine phosphorylation of Neu (p-Neu) in a dose-dependent manner (IC50~10 μg/ml) in tumor chunks, but had no effect on primary tumor epithelial cells grown in monolayer. Direct co-culture of mouse or human tumor epithelial cell lines with CAFs restored the epithelial cells' response to G129R, similar to that observed in mouse tumor chunks. The addition of PRL, as expected, induced p-Neu in both the tumor chunk and co-culture models. The inhibitory effect of G129R was absent when CAFs were physically separated from mouse tumor epithelial cells using a transwell system, or when CAFs were replaced with normal fibroblasts in direct co-culture with human or mouse tumor epithelial cells. In vivo, G129R reduced p-Neu levels in primary mammary tumors of mice in a time- and dose-dependent manner. In conclusion, CAFs play a critical role in bridging the cross-talk between PRL and HER2/Neu in both mouse and human models of breast cancer. The inhibitory effects of G129R on p-Neu and on tumor growth are dependent upon interactions of tumor epithelial cells with CAFs.     

Effects of cytokines derived from cancer-associated fibroblasts on androgen synthetic enzymes in estrogen receptor-negative breast carcinoma

Purpose

The tumor microenvironment plays pivotal roles in promotion of many malignancies. Cancer-associated fibroblasts (CAFs) have been well-known to promote proliferation, angiogenesis, and metastasis but mechanistic understanding of tumor–stroma interactions is not yet complete. Recently, estrogen synthetic enzymes were reported to be upregulated by co-culture with stromal cells in ER positive breast carcinoma (BC) but effects of co-culture on androgen metabolism have not been extensively examined. Therefore, we evaluated roles of CAFs on androgen metabolism in ER-negative AR-positive BC through co-culture with CAFs.

Methods

Concentrations of steroid hormone in supernatant of co-culture of MDA-MB-453 and primary CAFs were measured using GC–MS. Cytokines derived from CAFs were determined using Cytokine Array. Expressions of androgen synthetic enzymes were confirmed using RT-PCR and Western blotting. Correlations between CAFs and androgen synthetic enzymes were analyzed using triple-negative BC (TNBC) patient tissues by immunohistochemistry.

Results

CAFs were demonstrated to increase expressions and activities of 17βHSD2, 17βHSD5, and 5α-Reductase1. IL-6 and HGF that were selected as potential paracrine mediators using cytokine array induced 17βHSD2, 17βHSD5, and 5α-Reductase1 expression. Underlying mechanisms of IL-6 paracrine regulation of 17βHSD2 and 17βHSD5 could be partially dependent on phosphorylated STAT3, while phosphorylated ERK could be involved in HGF-mediated 5α-Reductase1 induction. α-SMA status was also demonstrated to be significantly correlated with 17βHSD2 and 17βHSD5 status in TNBC tissues, especially AR-positive cases.

Conclusions

Results of our present study suggest that both IL-6 and HGF derived from CAFs could contribute to the intratumoral androgen metabolism in ER-negative BC patients.

     

槲皮素通过调控p21/cip1和p27/kip1蛋白的稳定性抑制口腔鳞状细胞癌细胞增殖

目的:探究槲皮素通过调控p21/cip1和p27/kip1蛋白的稳定性对口腔鳞状细胞癌细胞增殖的影响.方法:CCK-8试剂盒检测口腔鳞状细胞癌细胞(SCC-15)增殖情况;流式细胞术检测SCC-15细胞周期分布;Western blot检测CyclinD1/CDK复合体、p21/cip1、p27/kip1、p-GSK3...     

Hepatocyte growth factor activates tumor stromal fibroblasts to promote tumorigenesis in gastric cancer

Cancer-associated fibroblasts (CAFs), as the activated fibroblasts in tumor stroma, are important modifiers of tumor progression. However, the mechanisms underlying stromal fibroblast activation and their promotion of tumor growth remain largely unknown in gastric cancer. Here, we show that normal fibroblasts (NFs) from non-cancerous regions of gastric cancer exhibit the traits of CAFs when grown together with gastric cancer cells in vivo. Activation of NFs can be induced by co-culture with gastric cancer cells, while deprivation of hepatocyte growth factor (HGF) using a neutralizing antibody inhibits the activation of NFs. Moreover, we identify HGF as an important factor from CAFs that acts in a paracrine manner to promote tumorigenesis in vitro and in vivo. Taken together, these results suggest that HGF may play a pivotal role in the regulatory circuit between gastric cancer cells and stromal fibroblasts, and neutralization of HGF inhibits both activation and tumor-promoting properties of CAFs.     

不同分化程度口腔鳞癌旁的成纤维细胞增殖活力比较

背景与目的：有关口腔鳞状细胞癌（OSCC）发生发展的机制目前尚未阐明,过去多数研究仅单纯从上皮的角度进行探讨,忽略了问质（宿主）细胞的变异在OSCC发生发展中的重要作用.本研究探讨在不同分化程度OSCC旁的癌相关成纤维细胞（CAFs）其增殖活性是否存在差异.方法：将前期冻存的第三代不同分化程度OSCC旁的CAFs和同部位正常成纤维细胞（NFs）进行复苏并鉴定,测定细胞生长曲线、有丝分裂指数曲线、MTT法,了解各类细胞的增殖活性是否存在差异.重复试验3次,每次检测3孔细胞,将平均值作为最终结果,并用两因素方差分析对检测值进行统计分析.结果：CAFs每天的细胞生长计数值、有丝分裂计数值、MTF检测值均高于NFs,CAFs的生长及存活能力均强于NFs（P＜0.05）,不同分化程度OSCC旁的CAFs增殖活性的差异有显著性（P＜0.05）,分化程度较低的舌癌旁CAFs的增殖活性更强.结论：OSCC与CAFs存在交互作用,CAFs增殖活性的改变可能对OSCC的分化程度产生影响.     

Altered TGF-β signaling in a subpopulation of human stromal cells promotes prostatic carcinogenesis

Carcinoma-associated fibroblasts (CAF) play a critical role in malignant progression. Loss of TGF-β receptor II (TGFβR2) in the prostate stroma is correlated with prostatic tumorigenesis. To determine the mechanisms by which stromal heterogeneity because of loss of TGFβR2 might contribute to cancer progression, we attenuated transforming growth factor beta (TGF-β) signaling in a subpopulation of immortalized human prostate fibroblasts in a model of tumor progression. In a tissue recombination model, loss of TGFβR2 function in 50% of the stromal cell population resulted in malignant transformation of the nontumorigenic human prostate epithelial cell line BPH1. Mixing fibroblasts expressing the empty vector and dominant negative TGFβR2 increased the expression of markers of myofibroblast differentiation [coexpression of vimentin and alpha smooth muscle actin (αSMA)] through elevation of TGF-β1 and activation of the Akt pathway. In combination, these two populations of stromal cells recapitulated the tumor inductive activity of CAFs. TGFβR2 activity in mixed stromal cell populations cultured in vitro caused secretion of factors that are known to promote tumor progression, including TGF-β1, SDF1/CXCL12, and members of the fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) families. In vivo, tissue recombination of fibroblasts overexpressing TGF-β1 and SDF1/CXCL12 not only induced transformation of BPH1 cells, but also promoted a robust growth of highly invasive cells, similar to effects produced by CAFs. While the precise nature and/or origin of the particular stromal cell populations in vivo remain unknown, these findings strongly link heterogeneity in TGF-β signaling to tumor promotion by tumor stromal cells.     

Oral Squamous Cell Carcinoma-Derived ANGPTL3 Induces Cancer Associated Fibroblastic Phenotypes in Surrounding Fibroblasts

Objective: Angiopoietin-like proteins (ANGPTLs) have emerged as both important regulator of lipid and glucose metabolism as well as insulin sensitivity. In particular, ANGPTL3 activity is one of the most important factors in cancer growth and invasion. Although ANGPTL3 have been studied in OSCC, but the role of ANGPTL3 between OSCC and CAFs has yet to be clearly defined. Thus, this study aimed to investigate the roles of ANGPTL3 in the differentiation of CAFs. Methods: For our study, we used hTERT-hNOFs to replace CAFs by coculturing them with oral squamous cell carcinoma (OSCC) cells. We did a microarray dataset analysis to investigate what factors secreted from OSCC cells can induce cancer associated fibroblastic phenotype in surrounding fibroblasts. The secreted factors were confirmed by RT-PCR, real-time PCR, and Western blot. Result: ANGPTL3 has the most secreted factor derived from various oral cancer cells. To investigate the role of ANGPTL3 in CAFs, we treated rhANGPTL3 in hTERT-hNOFs. The fibroblasts showed an increase of tumor-promoting cytokines (IL-6 and IL-8) and myofibroblastic markers, such as α-SMA and FAP. Conclusion: In conclusion, our study reports the first evidence that ANGPTL3 plays a crucial role in tumor microenvironments by inducing CAF. Therefore, targeting ANGPTL3 may be promising treatment strategy for CAF-targeted therapy in CAF-rich tumors.     

Bone marrow-derived myofibroblasts contribute to the mesenchymal stem cell niche and promote tumor growth

Carcinoma-associated fibroblasts (CAFs) that express α-smooth muscle actin (αSMA) contribute to cancer progression, but their precise origin and role are unclear. Using mouse models of inflammation-induced gastric cancer, we show that at least 20% of CAFs originate from bone marrow (BM) and derive from mesenchymal stem cells (MSCs). αSMA+ myofibroblasts (MFs) are niche cells normally present in BM and increase markedly during cancer progression. MSC-derived CAFs that are recruited to the dysplastic stomach express IL-6, Wnt5α and BMP4, show DNA hypomethylation, and promote tumor growth. Moreover, CAFs are generated from MSCs and are recruited to the tumor in a TGF-β- and SDF-1α-dependent manner. Therefore, carcinogenesis involves expansion and relocation of BM-niche cells to the tumor to create a niche to sustain cancer progression.     

Effects of FasL Expression in Oral Squamous Cell Cancer

Purpose: To probe the role of FasL in cell apoptosis in oral squamous cell carcinomas (OSCCs). Methods:The expression of Fas/FasL was assessed in 10 cases of normal oral epithelium, 38 cases of OSCC and tumorinfiltrating lymphocytes (TIL), and 11 cases of metastatic lymph nodes by immunohistochemistry. Apoptosisof tumor cells and TIL was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick endlabeling assay (TUNEL). FasL-induction of T cell apoptosis was tested by co-culture assay in vitro with SCC-9and Jurkat T cells. Results: The 10 cases of normal oral epithelium all demonstrated extensive expression of Fas,the positive rate being largely down-regulated in OSCC (21/38) (P<0.05) compared to the normal (10/10). At thesame time, the positive rate of FasL significantly increased in OSCC (P<0.05) especially those with lymph nodemetastasis (P<0.05). The positive rates of Fas in well and middle differentiated OSCC were higher than thosein poor differentiated OSCC (P<0.05). The AI of tumor cells in Fas-positive OSCC was remarkably higher thanthat in Fas-negative OSCC (P<0.01), with a positive correlation between Fas expression and cell differentiationas well as apoptosis (r=0.68, P<0.01). The AI of tumor cells in FasL positive OSCC was remarkably lower thanthat in control while the AI of TIL was higher than in FasL negative OSCC (P< 0.05). The AI of tumor cellsreversely correlated with that of TIL (r = -0. 72, P<0.05). It was found that SCC-9 cells expressing functionalFasL could induce apoptosis of Jurkat cells as demonstrated by co-culture assays. As a conclusion, it is evidentthat OSCC cells expressing FasL can induce apoptosis in Fas-expressing T cells. Conclusions: In progression ofOSCC, expression of the Fas/FasL changes significantly. The results suggest that FasL is a mediator of immuneprivilege in OSCC and may serve as an marker for predicting malignant change in oral tissues.     

Hyaluronan synthase 2 expressed by cancer-associated fibroblasts promotes oral cancer invasion

Background

Hyaluronan synthases (HAS) control the biosynthesis of hyaluronan (HA) and critically modulate the tumor microenviroment. Cancer-associated fibroblasts (CAFs) affect the progression of a tumor by remolding the matrix. However, little is known about the role of HAS from CAFs in this process. This study aimed to determine the role of hyaluronan synthase 2 (HAS2) from CAFs in the progression of oral squamous cell carcinoma (OSCC) invasion.

Methods

HAS isoforms 1, 2, and 3 in paired sets of CAFs and normal fibroblasts (NFs) were examined by real-time PCR, and the expression of HAS2 and α-SMA in OSCC tissue sections was further evaluated using immunohistochemical staining. Furthermore, we used a conditioned culture medium model to evaluate the effects of HAS2 from CAFs on the invasion and epithelial-mesenchymal transition (EMT) of the oral cancer cells Cal27. Finally, we compared the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) between CAFs and NF, and between CAFs with or without HAS2 knockdown using an antibody array and western blotting.

Results

CAFs expressed higher levels of HAS2 than the paired NFs. HAS2 expression was consistent with α-SMA-positive myofibroblasts in the stroma of OSCC, and these were significantly correlated advanced clinical stages and cervical lymph node metastasis. Knocking down HAS2 with a specific siRNA or treatment with a HAS inhibitor markedly attenuated CAF-induced invasion and EMT of Cal27 cells. Higher MMP1 and lower TIMP1 levels were detected in the supernatants of CAFs relative to NFs. Knocking down HAS2 could decrease the expression of MMP1 and increase that of TIMP1 in CAFs.

Conclusions

HAS2 is one of the key regulators responsible for CAF-mediated OSCC progression and acts by modulating the balance of MMP1 and TIMP1.

     

TNF‐α enhances TGF‐β‐induced endothelial‐to‐mesenchymal transition via TGF‐β signal augmentation

The tumor microenvironment (TME) consists of various components including cancer cells, tumor vessels, cancer‐associated fibroblasts (CAFs), and inflammatory cells. These components interact with each other via various cytokines, which often induce tumor progression. Thus, a greater understanding of TME networks is crucial for the development of novel cancer therapies. Many cancer types express high levels of TGF‐β, which induces endothelial‐to‐mesenchymal transition (EndMT), leading to formation of CAFs. Although we previously reported that CAFs derived from EndMT promoted tumor formation, the molecular mechanisms underlying these interactions remain to be elucidated. Furthermore, tumor‐infiltrating inflammatory cells secrete various cytokines, including TNF‐α. However, the role of TNF‐α in TGF‐β‐induced EndMT has not been fully elucidated. Therefore, this study examined the effect of TNF‐α on TGF‐β‐induced EndMT in human endothelial cells (ECs). Various types of human ECs underwent EndMT in response to TGF‐β and TNF‐α, which was accompanied by increased and decreased expression of mesenchymal cell and EC markers, respectively. In addition, treatment of ECs with TGF‐β and TNF‐α exhibited sustained activation of Smad2/3 signals, which was presumably induced by elevated expression of TGF‐β type I receptor, TGF‐β2, activin A, and integrin αv, suggesting that TNF‐α enhanced TGF‐β‐induced EndMT by augmenting TGF‐β family signals. Furthermore, oral squamous cell carcinoma‐derived cells underwent epithelial‐to‐mesenchymal transition (EMT) in response to humoral factors produced by TGF‐β and TNF‐α‐cultured ECs. This EndMT‐driven EMT was blocked by inhibiting the action of TGF‐βs. Collectively, our findings suggest that TNF‐α enhances TGF‐β‐dependent EndMT, which contributes to tumor progression.     

CCNB1基因通过调控PI3K/Akt信号通路对口腔鳞癌细胞增殖、侵袭和迁移的影响

目的 探讨细胞周期蛋白B1(CCNB1)基因对口腔鳞癌细胞增殖、侵袭和迁移的影响及作用机制。方法 实时荧光定量PCR(qRT-PCR)检测人正常口腔上皮细胞系HOK和人口腔鳞癌细胞系SCC-15、SCC-4、Cal-27中CCNB1基因的表达量,将CCNB1 siRNA(si-CCNB1)和阴性对照(si-NC)转染至SCC-15细胞,同时设置空白对照(Control),qRT-PCR和Western blot检测各组SCC-15细胞中CCNB1的表达,MTT实验、Transwell实验和划痕实验分别检测沉默CCNB1对SCC-15细胞增殖、侵袭和迁移能力的影响。Western blot检测细胞中MMP-2、MMP-9、PI3K、Akt和p-Akt蛋白的表达。结果 CCNB1在口腔鳞癌细胞系中的表达显著高于正常口腔上皮细胞(P＜0.05),si-NC组SCC-15细胞中CCNB1 mRNA和蛋白的表达与Control组无统计学差异(P＞0.05);si-CCNB1组中CCNB1 mRNA和蛋白的表达明显低于si-NC组和Control组(P＜0.05),si-NC组SCC-15细胞增殖、侵袭和迁移能力及MMP-2、MMP-9、PI3K、Akt、p-Akt蛋白表达与Control组无统计学差异(P＞0.05);si-CCNB1组细胞增殖、侵袭和迁移能力及MMP-2、MMP-9、PI3K、p-Akt蛋白表达均明显低于si-NC组和Control组(P＜0.05),两组间Akt蛋白表达无统计学差异(P＞0.05)。结论 CCNB1在口腔鳞癌细胞系中呈高表达,沉默CCNB1基因能够抑制人口腔鳞癌SCC-15细胞增殖、侵袭和迁移,其作用机制可能与抑制PI3K/Akt信号通路的激活有关。     

Sonic hedgehog signaling promotes growth of oral squamous cell carcinoma cells associated with bone destruction

Sonic hedgehog (Shh) and its signaling have been identified in several human cancers, and increased levels of its expression appear to correlate with disease progression and metastasis. However, the role of Shh in bone destruction associated with oral squamous cell carcinomas, which frequently invade the maxilla or the mandible, is still unclear. In this study we show that the use of siRNA for Shh to block SHH secreted by SAS oral squamous cell carcinoma cells suppressed the tumor growth and tumor angiogenesis of subcutaneous SAS xenografts in vivo. Moreover, blockade of Shh in SAS cells decreased tumor growth and osteoclast number in a tibial metaphysis mouse model. Significantly, we clearly show that SHH stimulated osteoclast formation in a co-culture system consisting of murine bone stromal ST2 cells and murine CD11b(+) bone marrow cells. These findings suggest that Shh signaling is a potential target for the treatment of oral squamous cell carcinoma associated with bone destruction.     

Therapeutic targeting of oncogenic transforming growth factor-β1 signaling by antisense oligonucleotides in oral squamous cell carcinoma

The transforming growth factor-β1 (TGF-β1) signaling pathway is important in human oral squamous cell carcinoma (OSCC). Accordingly, the aims of this study were to evaluate the effect of antisense TGF-β1 oligonucleotides (ODNs) on OSCC in cell culture and in a xenograft model, as well as to evaluate any effects ODNs have on proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2 (MMP-2) expression in the xenograft model. We performed real-time cell electronic sensing (RT-CES) to determine the effect of antisense TGF-β1 ODNs on SCC-9?cell growth. To examine the in?vivo effect of antisense TGF-β1 ODN therapy, SCC-9?cells were grafted into nude mice. Antisense ODNs were injected into the mass daily. Tumor size, body weight and duration of survival were assessed daily. Specimens from the main mass were used for immunohistochemical staining to analyze PCNA and MMP-2 expression. In?vitro treatment with antisense TGF-β1 ODNs decreased TGF-β1 expression and growth of SCC-9?cells. In the xenograft model, the antisense TGF-β1 ODN group exhibited a significantly decreased tumor growth rate compared to the control, which received Dulbecco's modified Eagle's medium (DMEM) (P=0.022). However, mean survival time and body weights were not significantly different between the groups (P>0.05). Immunohistochemistry showed that tumors from animals that received antisense TGF-β1 ODNs had significantly lower expression levels of PCNA and MMP-2 compared to tumors from animals in the DMEM group (P<0.05). In conclusion, antisense TGF-β1 ODN therapy significantly inhibits tumor growth compared to controls, however, there are no significant differences between groups with respect to changes in body weight.