Xiaoaiping injection affects the invasionand metastasis of hepatocellular carcinomaby controlling AFP expression

Objective Xiaoaiping (XAP) is a traditional Chinese medicine that is a commonly used as an anticancerdrug in clinical practice owing to its high efficiency and low toxicity. Specifically, XAP can effectively inhibitthe growth of hepatocellular carcinoma (HCC). Alpha-fetoprotein (AFP) is a key HCC diagnostic marker andis closely related to certain malignant cytological behaviors of HCC. However, whether AFP expression andXAP treatment are related to the invasion and metastasis of HCC remains unclear. In the present study, weaimed to evaluate the effects and underlying mechanism of XAP on the invasion and metastasis of HCC..Methods Using a cell scratch assay, Transwell technology, and western blotting we detected the differentinvasion and metastatic abilities of Hep3B cells in XAP treatment and blank control groups. This allowedcomparison of the invasion and metastatic abilities of Hep3B cells with differing levels of AFP expression.AFP mRNA sequencing technology was used to analyze the mechanism of tumor invasion and metastasisassociated with AFP and XAP treatment.Results Cell invasion and metastasis abilities in the XAP group were significantly lower than those in thecontrol group (P < 0.05). Additionally, compared to the control group, the expression of AFP significantlydecreased after XAP treatment (P < 0.05). The ability of Hep3B cells to invade and metastasize waspromoted when AFP expression was up-regulated, whereas it was inhibited when AFP was silenced. XAPinjection and AFP regulate the invasion and metastatic ability of HCC by affecting matrix metalloproteinases(MMPs).Conclusion XAP injection inhibits the invasion and metastatic ability of HCC by influencing the expressionof AFP;additionally, this inhibition of AFP is achieved by affecting MMPs.

Regarding on the Fractional Mathematical Model of Tumour Invasion and Metastasis

In this paper,we analyze the behaviour of solution for the system exemplifying model of tumour invasion and metastasis by the help of q-homotopy analysis transform method(q-HATM)with the fractional operator.The analyzed model consists of a system of three nonlinear differential equations elucidating the activation and the migratory response of the degradation of the matrix,tumour cells and production of degradative enzymes by the tumour cells.The considered method is graceful amalgamations of q-homotopy analysis technique with Laplace transform(LT),and Caputo–Fabrizio(CF)fractional operator is hired in the present study.By using the fixed point theory,existence and uniqueness are demonstrated.To validate and present the effectiveness of the considered algorithm,we analyzed the considered system in terms of fractional order with time and space.The error analysis of the considered scheme is illustrated.The variations with small change time with respect to achieved results are effectively captured in plots.The obtained results confirm that the considered method is very efficient and highly methodical to analyze the behaviors of the system of fractional order differential equations.

INVESTIGATION OF THE RELATIONSHIP BETWEEN CELL PROLIFERATIVE ACTIVITY AND INVASION, METASTASIS AND PROGNOSIS OF GASTRIC CANCER

To evaluated Bromodeoxyuridine (BrdUrd) /DNA doubleparametric method in detection of gastric carcinoma and to analyze the relationships of cellular BrdUrd labeling indices(LI), G2/M-phase fraction(G2/MPE) and DNA content to the depth of invasion, lymphatic vessel invasion, lymphatic node metastasis, peritoneal dissemination and blood vessel invasion and prognosis. Methods: Fresh tumor samples from 60 cases were examined by BrdUrd/DNA doubleparametric flowcytometry. Propidium iodide(PI) was used as fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used as second antibody. Results: The values of BrdUrd LI in patients with serosa invasion was significantly higher than those without serosa invasion (P<0.01); there was statistical significance in 5-year survival rate between the two groups (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic vessel invasion than those without invasion (P<0.01); the patients with lymphatic vascular invasion carried a significantly poor prognosis (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic node metastasis than those without metastasis (P<0.01), there was a statistical significance in 5 years survival between these 2 groups. The incidence of lymphatic node metastasis was significantly higher in aneuploid carcinoma (P<0.05), and the patients with aneuploid carried a significantly poor prognosis (P<0.05). Patients with peritoneal dissemination had a significantly worse prognosis (P<0.01). G2/MPF values were significantly higher in patients with blood vessel invasion than those without invasion (P<0.01). Conclusion: Cellular BrdUrd LI, G2/MPF and DNA content are related to depth of invasion, lymphatic vessel invasion, peritoneal dissemination, blood vessel invasion and prognosis of gastric carcinoma.

Blockade of the deubiquitinating enzyme USP48 degrades oncogenic HMGA2 and inhibits colorectal cancer invasion and metastasis

HMGA2,a pivotal transcription factor,functions as a versatile regulator implicated in the progression of diverse aggressive malignancies.In this study,mass spectrometry was employed to identify ubiquitin-specific proteases that potentially interact with HMGA2,and USP48 was identified as a deubiquitinating enzyme of HMGA2.The enforced expression of USP48 significantly increased HMGA2 protein levels by inhibiting its degradation,while the deprivation of USP48 promoted HMGA2 degradation,thereby suppressing tumor invasion and metastasis.We discovered that USP48 undergoes SUMOylation at lysine 258,which enhances its binding affinity to HMGA2.Through subsequent phenotypic screening of small molecules,we identified DUB-IN-2 as a remarkably potent pharmacological inhibitor of USP48.Interestingly,the small-molecule inhibitor targeting USP48 induces destabilization of HMGA2.Clinically,upregulation of USP48 or HMGA2 in cancerous tissues is indicative of poor prognosis for patients with colorectal cancer(CRC).Collectively,our study not only elucidates the regulatory mechanism of DUBs involved in HMGA2 stability and validates USP48 as a potential therapeutic target for CRC,but also identifies DUB-IN-2 as a potent inhibitor of USP48 and a promising candidate for CRC treatment.

Tumor microenvironment: driving forces and potential therapeutic targets for breast cancer metastasis

Distant metastasis to specific target organs is responsible for over 90%of breast cancer?related deaths,but the underlying molecular mechanism is unclear.Mounting evidence suggests that the interplay between breast cancer cells and the target organ microenvironment is the key determinant of organ?specific metastasis of this lethal disease.Here,we highlight new findings and concepts concerning the emerging role of the tumor microenvironment in breast cancer metastasis;we also discuss potential therapeutic intervention strategies aimed at targeting compo?nents of the tumor microenvironment.

Study on the mechanism of tRNA-ValAAC-5 in promoting the proliferation and migration of hepatocellular carcinoma cells

Objective:To demonstrate the role and mechanism of tRNA-ValAAC-5 expression in hepatocellular carcinoma(HCC)cells.Methods:The expression levels of tRNA-ValAAC-5 in HCC(Hep3B,HuH7,SNU398,Hep3G2)and human hepatocellular carcinoma(THLE2,THLE3)were detected by real-time PCR.HEP3B and Hep3G2 cells were respectively transfected with tRNA-ValAAC-5-inhibitor and tRNA-ValAAC-5-NC as the inhibitor group and the NC group.Then the ability of cell proliferation was detected by CCK-8 assay and the ability of invasion and metastasis was detected by Transwell assay.The protein expression levels of p21,Matrix metalloproteinase 2(MMP2)and Matrix metalloproteinase 9(MMP9)were determined by Western blot.Results:The relative expression of tRNA-ValAAC-5 in Hep3B,HuH7,SNU398 and Hep3G2 cells were significantly higher than THLE2 and THLE3 cells,the differences were statistically significant(P<0.05).After tRNA-ValAAC-5-inhibitor transfection,the expression of tRNA-ValAAC-5 in Hep3B and Hep3G2 cells were reduced than tRNA-ValAAC-NC group.Both of the differences were statistically significant(t=36.52,27.45,P<0.001),which indicated the transfection was successful.The proliferative ability of Hep3B and Hep3G2 cells transfected with tRNA-ValAAC-5-inhibitor after 24,48,72,96 h were inhibited effectively compared with tRNA-ValAAC-5-NC group.All of the differences were statistically significant in Hep3B(t=5.25,8.23,7.33,14.16,P<0.001)and Hep3G2(t=4.25,5.11,9.39,7.59,P<0.001)cells.The number of invasion and metastasis of Hep3B and Hep3G2 cells were reduced in tRNA-ValAAC-5-inhibitor group compared with tRNA-ValAAC-5-NC group,there was significant difference(t=14.01,21.85,P<0.001).The protein expression levels of P21 were lower,MMP2 and MMP9 were higher in tRNA-ValAAC-5-inhibitor group compared with tRNA-ValAAC-5-NC group,the differences were statistically significant in Hep3B(t=8.96,12.80,4.652,P<0.001)cells and Hep3G2(t=15.17,22.36,12.61,P<0.001)cells.Conclusion:tRNA-ValAAC-5 can effectively promote the proliferation,invasion and metastasis of HCC,and its possible mechanism is related to regulating the expression of p21,MMP2 and MMP9.

Role of tissue factor in hepatocellular carcinoma genesis, invasion and metastasis

Background Numerous studies indicate that tissue factor （TF）, namely tissue thromboplastin, has a close relationship with malignant tumor genesis and progress. It contributes to blood coagulation as well as the regulation of cellular differentiation, the formation of blood vessels, and also tumor recurrence and metastasis. The present study aimed to detect TF expression in hepatocellular carcinoma （HCC） patients and to elucidate its association with prognosis and clinical features of the disease. Methods The plasma TF levels of 50 HCC patients and 30 controls were assayed by ELISA. The expressions of TF mRNA and protein in HCC tissues, adjacent tissues and normal tissues were detected by reverse transcription- polymerase chain reaction （RT-PCR） and Western blotting. The acquired data were analyzed with related clinic-pathological documents. The patients were followed up for five years, and the relationship between TF and prognosis was analyzed. Results The plasma TF levels were significantly increased in HCC compared to the controls （P 〈0.05）, presenting a close relationship with differentiation level, tumor size and hepatocirrhosis occurrence （P 〈0.05）. There were remarkably higher values in cases of lymphatic metastasis, extrahepatic metastasis and portal tumor thrombus （PTT） （P 〈0.05） compared to non-metastasis or non-tumor thrombus, but no significant difference with different focus number or envelope （P 〉0.05）. The positive rates and the relative expression of TF mRNA in HCC tissue were 63.0% （17/27） and 0.567±0.268, respectively, significantly higher than that in adjacent tissues or normal tissues （P 〈0.05）. In the patients with positive results, the relative expression intensity varied significantly with different tumor size and index of local invasion and metastasis （P 〈0.05）. The positive rates and the relative expression intensities of TF protein in HCC tissue were 74.1% （20/27） and 4.093±1.256, respectively, significantly higher than those in adjacent tissue or normal tissue （P 〈0.05）. In the patients with positive results, the relative expression intensity showed significant difference in different tumor size, differentiation level, and index of local invasion and metastasis （P 〈0.05）. Conclusions The TF levels were significantly higher in plasma and tissues of HCC patients, presenting a close relationship with the index of invasion and metastasis. It indicated that TF might be related to differentiation and metastasis of HCC.

Positive association of RhoC gene overexpression with tumour invasion and lymphatic metastasis in gastric carcinoma

Worldwide estimates establish gastric carcinoma as the second most frequent cause of cancer deaths. Tumour invasion and metastasis is the biggest impediment to gastric carcinoma cure. Active migration of tumour cells is now considered as the pivotal step in cancer invasion and metastasis. RhoC is a member of the Ras-superfamily of small guanosine triphosphatases that can regulate many cellular functions, especially cytoskeletal organization and cell locomotion. Overexpressing RhoC in vitro in the poorly metastatic cell line from human melanoma may induce a highly metastatic phenotype.~1 The recent development of laser capture microdissection (LCM) affords the opportunity to further evaluate the role RhoC plays in the invasion and metastasis of gastric carcinoma cells in their native tissue environment.

Adverse effects of chemoradiotherapy on invasion and metastasis of tumor cells

The phenomenon of enhanced invasion and metastasis of residual tumor cells has been observed in an increasing number of patients receiving chemoradiotherapy recently,and tumor metastasis will undoubtedly limit patient prognosis.However,the key mechanism by which chemoradiotherapy affects the invasion and metastasis of tumor cells remains unclear.Studies have shown that chemoradiotherapy may directly act on tumor cells and alter the tumor microenvironment,or induce cell apoptosis and autophagy to promote tumor cell survival and metastasis.In this review,we summarize the potential mechanisms by which chemoradiotherapy may affect the biological behavior of tumor cells and open up new avenues for reducing tumor recurrence and metastasis after treatment.These insights will improve the efficacy of chemoradiotherapy.

Anti-rheumatic Drug Iguratimod(T-614) Alleviates Cancer-induced Bone Destruction via Down-regulating Interleukin-6 Production in a Nuclear Factor-κB-dependent Manner

Cytokines are believed to be involved in a “vicious circle” of progressive interactions in bone metastasis. Iguratimod is a novel anti-rheumatic drug which is reported to have the capability of anti-cytokines. In this study, a rat model was constructed to investigate the effect of iguratimod on bone metastasis and it was found that iguratimod alleviated cancer-induced bone destruction. To further explore whether an anti-tumor activity of iguratimod contributes to the effect of bone resorption suppression, two human breast cancer cell lines MDA-MB-231 and MCF-7 were studied. The effect of iguratimod on tumor proliferation was detected by CCK-8 assay and flow cytometry. The effects of iguratimod on migration and invasion of cancer cells were determined by wound-healing and Transwell assays. Results showed that high dose（30 μg/m L） iguratimod slightly suppressed the proliferation of cancer cells but failed to inhibit their migration and invasion capacity. Interestingly, iguratimod decreased the transcription level of IL-6 in MDA-MB-231 cells in a concentration-dependent manner. Moreover, iguratimod partially impaired NF-κB signaling by suppressing the phosphorylation of NF-κB p65 subunit. Our findings indicated that iguratimod may alleviate bone destruction by partially decreasing the expression of IL-6 in an NF-κB-dependent manner, while it has little effect on the tumor proliferation and invasion.

STUDIES ON THE GAP JUNCTIONAL INTERCELLULARCOMMUNICATION OF HUMAN NASOPHARYNGEALCARCINOMA CELLS AND THE EFFECT OF RII

Human nasopharyngeal carcinoma(NPC) cell line,CNE-2Z, and its clones(L2, H2, L4) with various invasive and metastatic potentials were examined for their gap junctions(GJ), gap junctional intercellular communication(GJIC) and the concentration of cytosolic free calcium(Ca2+). Only a few intermediate junction(IJ)but no GJ structures were observed under electron microscope(EM). CNE-2Z cells showed marked JGIC,while its variants lacked this function using the scraploading dye-transfer technique(SLDT). There was lower concentration of[Ca2+]. in L2 cells(a variant with high invasive and metastatic Potential) compared to that in H2 and L4 cells(variants with medium and low invasive and metastatic Potentials, respectively). These data suggested that high invasive and metastatic potentials might be correlated with the levcl of[Ca2+]i in NPC cells.The effect of RII(4-hydroxycarbophenyl retinamide) on NPC cells also investigated, After 3-7 d of RII(10-5 M) treatment, there was no change in the number of gap junctions and other kind of intercellular junctions in NPC cells observed under EM. The JGIC of CNE-2Z weaked and then disappeared finally with prolonging of RII treatment. However. there was no influence on its variants. The level of[Ca2+], in NPC cells apparently fell after 6 h of RII treatment, and rose to original level with persisting of RII treatment. Whether the fluctuating of[Ca2+]i level is related to the inhibitory effect of RII treatment on growth and invasion of NPC cells needs to be further studied.

Role of adhesion molecules in cancer and targeted therapy

Adhesion molecules mediate cell-to-cell and cell-to-extracellular matrix interactions and transmit mechanical and chemical signals among them.Various mechanisms deregulate adhesion molecules in cancer,enabling tumor cells to proliferate without restraint,invade through tissue boundaries,escape from immune surveillance,and survive in the tumor microenvironment.Recent studies have revealed that adhesion molecules also drive angiogenesis,reshape metabolism,and are involved in stem cell self-renewal.In this review,we summarize the functions and mechanisms of adhesion molecules in cancer and the tumor microenvironment,as well as the therapeutic strategies targeting adhesion molecules.These studies have implications for furthering our understanding of adhesion molecules in cancer and providing a paradigm for exploring novel therapeutic approaches.

Association of E-cadherin and β-catenin with metastasis in nasopharyngeal carcinoma

Background This study was designed to detect methylation of E-cadherin gene promoter and gene mutation of β-catenin in exon 3 and their expression of protein and mRNA in primary tumor and lymph node metastatic tumor of nasopharyngeal carcinoma (NPC), and investigate the mechanism of invasion and metastasis of neoplastic cells in NPC Methods Fourty-two fresh biopsy samples were taken from untreated NPC patients at the Affiliated Hospital of Sun Yat-sen Medical College, Sun Yat-sen University, Guangzhou, China during the period of 1999-2002 Among them 21 were taken from primary tumors and the other 21 from lymph node metastatic tumors The gene promoter methylation of E-cadherin was detected by methylation-specific PCR (MSP) The mutation in exon 3 of β-catenin was detected by direct sequencing analysis RT-PCR, Western blot and immunohistochemical staining were used to detect the mRNA and protein expression patterns in both primary and metastatic tumors of NPC Results Down-regulated expression of E-cadherin in metastatic tumor was compared with that in primary tumor Reduced expression of E-cadherin was found to be correlated with lymph node metastatic tumor of NPC ( P =0 004); but there was no obvious correlation between primary and metastatic tumors in the expression of β-catenin ( P =0 698) The mRNA expression level of E-cadherin in metastatic tumors decreased significantly compared with that in primary tumors However, little change was observed in the mRNA level of β-catenin in different tumor tissues Only 4 samples (19 1%) displayed gene promoter methylation of E-cadherin in primary tumor and 10 samples (47 6%) showed methylated form of E-cadherin The gene promoter methylation of E-cadherin was more common in metastatic tumor than in primary tumor of NPC ( P =0 024) Only 2 (4 76%) of the 42 samples showed mutations in exon 3 of β-catenin at 41 (T41A, ACCGCC) and codon 47 (S47T, AGTACT) The cytoplasmic and nuclear expression of β-catenin in tumor was not found in any samples of NPC Conclusions The results suggest that the downregulation of E-cadherin results from the gene promoter aberrant methylation of E-cadherin and that the methylation of E-cadherin plays an important role in invasion and metastasis of tumor cells in NPC However, β-catenin mutation is an infrequent event in NPC, and β-catenin is not a critical factor influencing the invasion and metastasis of tumor cells in NPC

Relations of proliferative activities of gastric carcinoma cells to lymphatic involvement, venous invasion and prognosis

Background This study was to evaluate bivariate bromodeoxyuridine(BrdUrd)/DNA flow cytometric analysis in detection of gastric carcinoma and to study the relations of cellular BrdUrd labeling indices (LI), G_2/M-phase fraction(G_2/MPF) and DNA ploidy pattern to lymphatic involvement, venous invasion and prognosis.Methods Fresh tumor samples from 60 patients with gastric carcinoma were analyzed by bivariate BrdUrd/DNA flow cytometry. The results were correlated with lymphatic vessel invasion, lymphatic node metastasis, the number of matastatic lymphatic nodes, and venous invasion. Propidium iodide (PI) was used as a fluorescent probe for total cellular DNA, and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescent-labeled goat anti-mouse antibody was used as a second antibody. S-phase fractions were measured by in vitro BrdUrd labeling, and DNA ploidy and G_2/MPF were also measured. Comparison of survival was performed with the log-rank test, the Chi-square test for qualitative data, and Student’s t test for quantu data. Results BrdUrd LI and G_2/MPF values were significantly higher in tumors with lymphatic vessel invasion than in those without invasion respectively (P<0.01); the patients who had tumors with lymphatic vessel invasion showed a significantly poor prognosis (P<0.01). Both BrdUrd LI and G_2/MPF values were significantly higher in tumors with lymphatic node metastasis than in those without metastasis (P<0.01). A statistical significant difference was noted in the 5-year survival rates between the patients with lymph node metastasis and those without metastasis. Compared with diploid carcinoma, the incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P<0.05), and the patients with aneuploid carcinoma showed a significantly poor prognosis (P<0.05). BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1-4 metastatic lymph nodes (P<0.05) and those without metastasis (P<0.01). G_2/MPF values in those patients either with more than 5 metastatic lymph nodes or 1-4 metastatic lymph nodes were higher than those without metastasis (P<0.01 and P<0.05). A statistical significance was seen in the 5-year survival rates among the patients with no metastatic lymph node, 1-4 metastatic nodes and more than 5 metastatic nodes (P<0.01). G_2/MPF values were significantly higher in patients with venous invasion than in those without invasion (P<0.01).Conclusions Positive correlations exist between cellular BrdUrd LI, G_2/MPF with lymphatic involvement and prognosis, and DNA aneuploid with lymphatic involvement and prognosis. The same was true between G_2/MPF value and venous invasion in gastric carcinoma.