Reciprocal regulation of RhoA and RhoC characterizes the EMT and identifies RhoC as a prognostic marker of colon carcinoma

Understanding how RhoC expression and activation are regulated is essential for deciphering its contribution to tumorigenesis. Here, we report that RhoC expression and activation are induced by the epithelial to mesenchymal transition (EMT) of colon carcinoma. Using LIM 1863 colon cancer cells, RhoC protein expression and subsequent activation were detected coincident with the loss of E-cadherin and acquisition of mesenchymal characteristics. Several Ets-1 binding sites were identified in the RhoC promoter, and evidence was obtained using chromatin immunoprecipitation that Ets-1 can regulate RhoC expression during the EMT. Interestingly, a marked decrease in RhoA activation associated with the EMT was observed that corresponds to the increase in RhoC expression. Use of shRNA established that RhoA inhibits and RhoC promotes post-EMT cell migration, demonstrating functional significance for their coordinate regulation. To assess the importance of RhoC expression in colon cancer, immunohistochemistry was performed on 566 colorectal tumors with known clinical outcome. The level of RhoC ranged from no expression to high expression, and statistical analysis revealed that elevated RhoC expression correlates with poor outcome as well as aberrant expression and localization of E-cadherin. These data provide one mechanism for how RhoC expression is regulated in colon carcinoma and substantiate its utility as a prognostic marker.     

Notch1 regulates the functional contribution of RhoC to cervical carcinoma progression

Background:

The role of Notch signalling in human epithelial cancers is of immense interest. In this study, we examine the interplay between Notch signalling and RhoC, a well-established molecular factor in metastasis. By linking the function of Notch and RhoC, we further strengthen the notion that there is a pro-oncogenic role of Notch signalling in human cervical cancers.

Methods:

RhoC protein expression in cervical carcinoma cell lines was assessed by western blotting. Using CaSki and SiHa cells (cervical carcinoma cells lines), we show that RhoC contributes to wound healing, invasion and migration, anoikis resistance, colony formation, in vitro tube formation and tumour formation. Immunohistochemical studies were carried out to assess the co-expression of RhoC, pAkt and Notch1 in clinical sections.

Results:

An assessment of the changes associated with epithelial-to-mesenchymal transition (EMT) shows that both Notch1 and RhoC have similar phenotypic contribution to EMT. Rho activity assessment on Notch1 inhibition with DAPT shows decreased RhoC activity. We further show that constitutively active RhoC rescues the phenotypic effect of Notch1 inactivation, and a comparison of Notch1 with RhoC expression shows an overlap between the two proteins in the same areas of the tissue.

Conclusion:

This study has provided evidence to suggest that RhoC is an effector of Notch1 in cervical carcinoma.     

Targeted disruption of protein kinase C epsilon reduces cell invasion and motility through inactivation of RhoA and RhoC GTPases in head and neck squamous cell carcinoma

Over 70% of patients with head and neck squamous cell carcinoma (HNSCC) present with locoregionally advanced stage III and IV disease. In spite of aggressive therapy, locoregional disease recurs in 60% and metastatic disease develops in 15% to 25% of patients causing a major decline in quality and length of life. Therefore, there is a need to identify and understand genes that are responsible for inducing an aggressive HNSCC phenotype. Evidence has shown that protein kinase C (PKC) epsilon is a transforming oncogene and may play a role in HNSCC progression. In this study, we determine the downstream signaling pathway mediated by PKC epsilon to promote an aggressive HNSCC phenotype. RNA interference knockdown of PKC epsilon in UMSCC11A and UMSCC36, two highly invasive and motile HNSCC cell lines with elevated endogenous PKC epsilon levels, resulted in cells that were significantly less invasive and motile than the small interfering RNA-scrambled control transfectants; 51 +/- 5% (P < 0.006) and 49 +/- 3% (P < 0.010) inhibition in invasion and 69 +/- 1% (P < 0.0005) and 66 +/- 3% (P < 0.0001) inhibition in motility, respectively. PKC epsilon-deficient UMSCC11A clones had reduced levels of active and serine-phosphorylated RhoA and RhoC. Moreover, constitutive active RhoA completely rescued the invasion and motility defect, whereas constitutive active RhoC completely rescued the invasion and partially rescued the motility defect of PKC epsilon-deficient UMSCC11A clones. These results indicate that RhoA and RhoC are downstream of PKC epsilon and critical for PKC epsilon-mediated cell invasion and motility. Our study shows, for the first time, that PKC epsilon is involved in a coordinated regulation of RhoA and RhoC activation, possibly through direct post-translational phosphorylation.     

乳腺浸润性微乳头状癌临床病理分析

目的:探讨乳腺浸润性微乳头状癌(IMPC)的临床病理特征、诊断、鉴别诊断及预后。方法:回顾性分析7例乳腺IMPC患者的临床表现、组织病理特点、免疫组化结果,同时进行文献复习。结果:7例乳腺IMPC患者无特殊临床症状和体征。组织病理特点:IMPC肿瘤细胞团排列形成不含纤维血管轴心的微小乳头状或假腺管状结构,其外侧缘呈锯齿或毛刺状;每个肿瘤细胞团与周边的间质之间存在大小不等的腔隙,其内缺乏内容物。免疫组化染色EMA阳性部位在癌细胞巢团的外缘。结论:乳腺IMPC是一种特殊类型的乳腺浸润性癌,比较少见,但因其具有高淋巴血管侵犯、高淋巴结转移等恶性生物学行为,预后较差,应引起病理及临床医生的足够重视。     

D2-40与乳腺浸润性导管癌临床病理分析

目的:观察淋巴管密度与乳腺非特殊类型浸润性导管癌(IDC,NOS)临床病理关系。方法:用淋巴管内皮细胞抗体D2-40单克隆抗体标记70例乳腺IDC(NOS)淋巴管表达,计算淋巴管密度,并同时进行ER、PR、HER-2、p53、VEGF和CD44V6检测,分析LVD与各临床病理指标的关系。结果:在IDC(NOS)中,D2-40标记的LVD与临床分期和腋淋巴结转移有关(P<0.05),而与年龄、组织学分级、及肿瘤直径无关(P>0.05)。进一步对临床分期行L-S-D检验,发现临床分期I期中的LVD与临床分期II、III期中的LVD有显著性差异,而临床分期II、III期间的LVD并无显著性差异。在IDC(NOS)中,D2-40标记的LVD与CD44V6表达相关(P<0.05),与ER、PR、HER-2、p53及VEGF的表达无关(P>0.05)。多因素分析显示,在IDC(NOS)中,D2-40标记的LVD与腋淋巴结转移和CD44V6有关(P<0.05),而与年龄、组织学分级、肿瘤直径、临床分期、ER、PR、HER-2、p53及VEGF无关(P>0.05)。结论:在IDC(NOS)中,D2-40标记的LVD与淋巴结状态相关,提示LVD可能与乳腺癌的预后有关,可能成为乳腺癌预后的独立指标。在IDC(NOS)中,D2-40标记的LVD与CD44V6表达呈正相关,并具有统计学意义,提示在肿瘤的生长过程中,CD44V6可能通过裂解细胞间质,促进肿瘤生长,导致间质中淋巴管密度增加,提高了肿瘤转移的机率。     

Farnesyl transferase inhibitor treatment of breast cancer cells leads to altered RhoA and RhoC GTPase activity and induces a dormant phenotype

Farnesyl transferase inhibitors (FTIs) were shown to be effective in modulating tumor growth in Ras‐transformed tumor cells. Recent studies have focused on Rho GTPases as putative targets of FTI action. Previously, we demonstrated that FTIs were effective in inhibiting the growth and invasiveness of RhoC GTPase‐overexpressing inflammatory breast cancer (IBC) cells however, RhoC activity was increased. In this study, we examine the mechanisms of FTI action on breast cancer cells in culture through modulation of RhoC and RhoA GTPases. We found that FTI inhibition of breast cancer cell growth was reversible and resembled what has been described for an in vitro model of tumor cell dormancy. On FTI treatment, levels of active RhoA decreased significantly, whereas levels of active RhoC increased 3.8‐fold. We studied the role of these two GTPases in a fibronectin and basic FGF‐induced model of breast cancer cell dormancy. Hypoactivation of RhoA and hyperactivation of RhoC were seen to induce morphology and growth changes consistent with tumor cell dormancy in culture. In addition, the JNK/SAPK pathway was induced on FTI treatment. A pharmacologic inhibitor of the JNK/SAPK pathway significantly reduced the number of dormant cells. This study has implications for the use of FTIs as therapeutic agents as well as potential mechanisms for breast cancer cell dormancy.     

乳腺浸润性微乳头状癌104例临床与病理分析

目的探讨乳腺浸润性微乳头状癌（invasive micropapillary carcinoma,IMPC）的临床病理特征、诊断、综合治疗和预后。方法对104例IMPC患者进行回顾性分析。结果104例IMPC无特殊的症状和体征,占同期全部乳腺癌9364例的1.1%。其淋巴结转移率（77.0%）及转移个数（平均9.7枚）均明显高于浸润性导管癌（41.0%,平均3.5枚）,二者比较有统计学意义（P〈0.05）。随访至2009年7月初,生存率3年75.0%、2年85.2%、1年96.3%;无瘤生存率3年70.8%、2年81.5%、1年95.0%。结论乳腺IMPC是一种近几年才受到重视的乳腺癌类型,具有不可忽视的发病率,以及淋巴管侵袭力强、淋巴结转移率高、预后差的生物学行为。     

Early invasive mucinous carcinoma of the breast

Cases of early invasive mucinous carcinoma of the breast have been reviewed. Microscopically, a typical lesion exhibits leakage of cancerous mucus into the stromal tissue from an intraductal carcinomatous component which were shown to be mucus-producing and to be lower papillary projection. Among 5,360 primary breast cancers that had been mastectomized at the C.I.H., 11 or 0.21% were determined as being "early" cases. All of these cases exhibited "pure type" mucinous carcinomas which were found to be highly mucus. The average age of these patients was 41 years (range: 34-51 yrs.), and no cases showed a nodal involvement. During an average follow-up of 8 years, no recurrent case were found. Thus, we have found that these "early" cases revealed an early invasion of a mucinous carcinoma that is highly mucus-producing but without an invasive ductal carcinomatous pattern.     

Regulation of in situ to invasive breast carcinoma transition

The transition of ductal carcinoma in situ (DCIS) to invasive carcinoma is a poorly understood key event in breast tumor progression. Here, we analyzed the role of myoepithelial cells and fibroblasts in the progression of in situ carcinomas using a model of human DCIS and primary breast tumors. Progression to invasion was promoted by fibroblasts and inhibited by normal myoepithelial cells. Molecular profiles of isolated luminal epithelial and myoepithelial cells identified an intricate interaction network involving TGFbeta, Hedgehog, cell adhesion, and p63 required for myoepithelial cell differentiation, the elimination of which resulted in loss of myoepithelial cells and progression to invasion.     

The classification of invasive carcinoma of the breast

The classification of invasive breast carcinoma assists diagnosis, allows for comparison of different patient groups in clinical trials and facilitates epidemiological analysis. For the individual patient, accurate tumor classification informs clinical decision-making with emphasis on assessment of prognosis and treatment formulation. Tumor grade is an independent prognostic indicator and is calculated by assessing specific tumor characteristics microscopically. The Tumor Node Metastasis staging system, produced by the American Joint Committee on Cancer Union for International Cancer Control, combines information about the primary tumor size, the status of the regional lymph nodes and the presence or absence of distant metastases at diagnosis to classify disease. In recent years, the use of gene expression profiling technology has led to the development of the molecular classification of breast cancer and has highlighted the importance of hormone receptor and HER2 oncogenic pathways, with particular reference to targeted chemotherapy. Tumor typing involves the identification of ‘no special type’ carcinoma with variable clinical, histological and molecular characteristics and ‘special type’ carcinomas that are usually associated with a particular set of prognostic and predictive indices. Some special type carcinomas have unique biological features that influence diagnostic investigation and clinical management.     

Different responses to preoperative chemotherapy for invasive lobular and invasive ductal breast carcinoma.

AIM: Preoperative chemotherapy (PCT) is used in primary breast cancer, to facilitate breast conservative surgery (BCS). Clinical and pathologic responses are important prognostic parameters. Biologic markers are needed to individualize treatment. PATIENTS AND METHODS: One hundred and thirty-five patients with breast carcinoma were treated with PCT, followed by surgery and adjuvant therapy. Clinical response and pathological complete response (pCR), biological markers and type of surgery were compared between invasive ductal (IDC) and invasive lobular carcinoma (ILC). RESULTS: Overall response (OR) for IDC was 75% compared to 50% for ILC (P=0.0151). Pathological CR was 15% for IDC and 0% for ILC (P=0.0066). Fifty-six percent of the responding patients had BCS, in contrast with 16% of the non-responders. BCS was performed in 50% of patients with IDC, in 38% of the patients with ILC. Salvage surgery was more necessary in ILC (19%) compared to IDC (4%) (P=0.0068). Patients with ILC were more frequently ER-positive and HER-2 negative than patients with IDC. CONCLUSIONS: Clinical and pathological responses are lower in ILC compared to IDC. After PCT, patients with large ILC should preferably be offered mastectomy with immediate breast reconstruction. However, PCT still remains valuable to evaluate tumor response and biologic factors in vivo.     

乳房派杰氏病合并浸润性乳腺癌 患者的生存分析

背景与目的:乳房派杰氏病(Paget's disease)是一种很罕见的疾病.文献报道该病患者的生存情况与其下乳房内的病灶的恶性程度相关.该研究旨在探索合并浸润性乳腺癌的乳房派杰氏病的临床病理特征并分析患者的生存率.方法:2002—2007年,共54例在复旦大学附属肿瘤医院接受治疗的合并浸润性乳腺癌的乳房派杰氏病患者纳入该研究.由于乳腺派杰氏病大多数为人表皮生长因子受体2(human epidermal growth factor receptor 2,HER-2)阳性,研究者随机收集了同期72例HER-2阳性浸润性乳腺癌患者作为对照组,比较两组的生存差异.结果:25例(46.3％)研究组患者没有典型的乳头乳晕派杰氏病皮肤表现,35例(64.8％)研究组患者浸润性癌灶超过2 cm,26例(48.1％)研究组患者腋窝淋巴结有转移.与对照组的患者相比,派杰氏病患者的复发事件更多(5年无复发生存率:76.4％vs 48.5％,P<0.01).对照组复发事件多集中于术后3年内,而派杰氏患者复发风险在术后5年内持续存在.研究组的5年总生存率(61.0％)也低于对照组(80.6％,P=0.01).结论:合并浸润性癌的乳房派杰氏病患者比HER-2阳性乳腺癌患者的预后差.     

乳腺浸润性导管癌预后相关因素分析

目的:探讨乳腺浸润性导管癌预后相关因素.方法: 收集130例乳腺浸润性导管癌资料,回顾性分析其临床特征、病理分化程度、复发转移情况、激素受体状况、人类表皮生长因子受体2的表达、临床治疗及生存情况.结果: c-erbB-2表达在ER、PR阳性组低于ER、PR阴性组(P<0.01),ER表达在PR阳性组高于PR阴性组(P<0.01);中、低分化与高分化相比,在淋巴结的转移、肿瘤的转移或复发、临床分期、肿块的大小上,均有统计学差异(P<0.05);单因素分析结果显示,激素受体状况、人类表皮生长因子受体2表达、病理分化程度、淋巴结状况、肿瘤转移或复发、临床分期、肿瘤大小、T分期、N分期、辅助化疗等11个因素与预后相关;多因素分析结果显示:ER状况、病理分化程度、淋巴结状况、临床分期是乳腺浸润性导管癌患者预后的独立影响因素.结论: 对乳腺浸润性导管癌,早期发现并针对病理分化程度及激素受体水平的适当治疗是提高生存期的关键.     

乳腺浸润性导管癌预后相关因素分析

目的：探讨乳腺浸润性导管癌预后相关因素。方法：收集130例乳腺浸润性导管癌资料,回顾性分析其临床特征、病理分化程度、复发转移情况、激素受体状况、人类表皮生长因子受体2的表达、临床治疗及生存情况。结果：c—erbB-2表达在ER、PR阳性组低于ER、PR阴性组（P〈0．01）,ER表达在PR阳性组高于PR阴性组（P〈0．01）;中、低分化与高分化相比,在淋巴结的转移、肿瘤的转移或复发、临床分期、肿块的大小上,均有统计学差异（P〈0．05）;单因素分析结果显示,激素受体状况、人类表皮生长因子受体2表达、病理分化程度、淋巴结状况、肿瘤转移或复发、临床分期、肿瘤大小、T分期、N分期、辅助化疗等11个因素与预后相关;多因素分析结果显示：ER状况、病理分化程度、淋巴结状况、临床分期是乳腺浸润性导管癌患者预后的独立影响因素。结论：对乳腺浸润性导管癌,早期发现并针对病理分化程度及激素受体水平的适当治疗是提高生存期的关键。     

乳腺浸润性微乳头状癌的临床病理特征及预后分析

目的:探讨乳腺浸润性微乳头状癌(invasive micropapillary carcinoma,IMPC)的临床病理特征及预后。方法:回顾性分析2011年1月至2015年12月246例于天津医科大学肿瘤医院收治的乳腺IMPC患者的临床病理资料,分为143例IMPC成分比例>50%(A组)和103例比例≤50%(B组)两组。多因素分析采用Cox比例风险回归模型,采用Log-rank检验及Kaplan Meier法等进行生存分析。结果:A组患者的5年无病生存(disease free survival,DFS)和总生存(overall survival,OS)时间均低于B组DFS(76.5%vs. 83.6%,P=0.042)和OS(74.1%vs. 81.6%,P=0.029)。A组中未行放疗患者的DFS和OS均低于行放疗患者的DFS(χ²=5.219,P=0.022)和OS(χ²=3.963,P=0.047)。Cox比例风险回归模型多因素分析显示,患者人类表皮生长因子受体-2(human epidermal growth facto...     

乳腺浸润性微小乳头状癌的诊断

淋巴结转移率高、预后差是乳腺浸润性微小乳头状癌的独特生物学行为.通过光镜下观察到肿瘤细胞聚集成微乳头状,以及免疫组化上皮膜抗原(EMA)强表达是准确及时诊断该类型肿瘤的可靠方法.相比较以往的诊断标准,最近提出的新标准更加重视乳腺浸润性微小乳头状癌的诊断,因而对这类患者的治疗及预后判断有重要的意义.     

乳腺浸润性微乳头状癌的研究现状

乳腺浸润性微乳头状癌（invasive micropapillary carcinom,IMPC）是乳腺癌一种特殊类型,具有独特的微乳头状结构及生长方式,发病率较低,约占所有乳腺癌的3%～6%。相比较于其他类型浸润性乳腺癌,IMPC具有更高的淋巴结转移率和更强的淋巴管侵袭性的特征,其恶性程度高、预后差,近年来引起国内外病理以及临床医师的高度关注。