Experimental Study On The Mechanism Of Vasculogenic Mimicry Formation In Hepatocellular Carcinoma

It is still a urgent problem that the metastasis and recurrence during currenthepatocellular carcinoma (HCC) treating. Patients with hepatocellular carcinoma(HCC) after hepatectomy occur metastasis and recurrence72.3%in5years, also34.5%in small HCC, thereinto,3/4in the first2years. However, it is not definite about themechanism of metastasis and recurrence. Research results mainly focus on: metastaticgene and non-metastatic gene in HCC, local invasion and motility, tumor cell adhesion,neo-vascularization, extracellular matrix, immune status and others. For the past fewyears, researchers have discovered the phenomenon in human highly aggressive uvealmelanoma, vasculogenic mimicry. Further clinical investigations present that it relateswith tumor blood supply and metastasis and recurrence, thus those point a new way totreat tumor metastasis and recurrence. Inhibiting neo-vascularization may be animportant method of inhibiting HCC cell growth, metastasis and recurrence for thereason that it shows tumor angiogenesis characteristic of affluent blood supply in HCC.In the research, the mechanisms of tumor vasculogenic mimicry mainly focus on: tumorcell itself, extracellular matrix, microenvironment and other aspects. According topublished reports, cancer stem cells participate in the formation of vasculogenicmimicry in acute leukemic and glioblastoma, however, whether in HCC has been fewreported. PART ⅠExpression and significance of endothelial related molecules in vasculogenicmimicry of hepatocellular carcinomaObjective: To explore the relationship between endothelial relatedmolecules(VE-cadherin and CD31) expression and vasculogenic mimicry ofhepatocellular carcinoma in vitro.Methods: We established three-dimensional cell cultures in several cell lines follow asHCC97H cell, SMMC7721cell and L02cell, and observed the ability to formvasculogenic mimicry. Interesting cells which formed vasculogenic mimicry wereisolated by the laser capture microdissection, and analyzed by semi-quantitative reversetranscription-polymerase chain reaction and Western blot subsequently.Results: HCC97H cell had the ability to form vasculogenic mimicry inthree-dimensional cell cultures, while SMMC7721and L02cell hadn’t the ability.Interesting cells had been captured by LCM successfully. In comparative analysis, theexpression level of VE-cadherin gene and protein in different groups were0.596±0.020、0.430±0.010and0.170±0.0100;1.536±0.120、1.176±0.055、0.780±0.060,respectively (P<0.05), whereas CD31was not expressed by any of the liver cancer celllines.Conclusions: Highly aggressive liver cancer cell has the ability to form vasculogenicmimicry in vitro, while poorly aggressive liver cancer cell and normal liver cellhaven’t. And VE-cadherin has impact on the formation of vasculogenic mimicry inHCC. PART ⅡExpression and significance of CD34and CD133in vasculogenic mimicry of livercancer cellsObjective: To investigate the relationship between expression of CD34and CD133andvasculogenic mimicry of hepatocellular carcinoma in vitro.Methods: We established three-dimensional cell cultures in several cell lines follow asHCC97H cell, SMMC7721cell and L02cell, and observed the ability to formvasculogenic mimicry. Interesting cells which formed vasculogenic mimicry wereisolated by the laser capture microdissection, and analyzed by semi-quantitative reversetranscription-polymerase chain reaction and Western blot subsequently.Results: HCC97H cell had the ability to form vasculogenic mimicry inthree-dimensional cell cultures, while SMMC7721and L02cell hadn’t the ability.Interesting cells have been captured by LCM successfully. In comparative analysis, thegene expression level of CD133and CD34in different groups were1.333±0.1001、0.9200±0.04583、0.3667±0.02517and0.8913±0.01935、0.6087±0.02173、0.1970±0.004(P<0.05); the protein expression level of CD133and CD34in differentgroups were0.8223±0.03009、0.4623±0.04382、0.2833±0.01582；0.775±0.0459、0.5617±0.02608、0.1903±0.06099(P<0.05), respectively.Conclusions: Highly aggressive liver cancer cell has the ability to form vasculogenicmimicry in vitro, while poorly aggressive liver cancer cell and normal liver cell haven’t. And cancer stem cells has impact on the formation of vasculogenic mimicry inHCC.