Studies On Metastatic Biological Characteristics Of Different Metastatic Potential Human Laryngeal Carcinoma Cell Subclones

Objective To isolate human laryngeal carcinoma cell subclones with different potential of metastasis.Methods High and low metastasis potential cell clones were isolated by improved invasion assay. Cell growth and cycle time were detected by cell growth curve assay, doubling time, and flow cytometry. Cell invasiveness and migration ability were analyzed by invasion assay and migration assay. Nude mice were used to study the growth and metastasis abilities in vivo.Results High and low metastasis potential cell clones were isolated successfully. Of the high and low metastasis potential cell clones, the doubling time were (31.6±1.1)h and (60.2±11.1)h(P<0.05), the proliferation index were (27.6±0.5)% and (16.3±0.8)% ( P < 0.01 )respectively ,the number of invasive cells were 210.2±12.9 and 11.6±5.0. (P<0.01),the number of migratory cells were 229.8±12.1 and 6.0±4.5(P<0.01). In vivo, the high metastasis potential cell clones group grew faster than the low metastasis potential cell clones group, with 2 weeks and 4 weeks to maturate respectively. The pathological exam showed metastasis in 3 ipsilateral popliteal lymph node, 4 ipsilateral inguinal lymph node and 1 contralateral inguinal lymph node in high metastatic group; 2 ipsilateral inguinal lymph node in low metastatic group. Reactive hyperplasia was detected in 1 ipsilateral inguinal lymph node in low metastatic group. In the high metastasis potential group, the weight of tumor was (17.0±3.0) mg, and the vascular count in the tumor was 21.2±4.0. In the low metastasis potential group, the weight of tumor was ( 9.7±3.8) mg(P<0.05) and the vascular count in the tumor was 1.4±0.9(P<0.01). There were more VEGF-C mRNA and protein in the high metastasis potential group then in the low metastasis potential group ( P<0.05) .Conclusion We successfully isolated the high and low metastasis potential cell clones from one cell line of laryngeal carcinoma. The high metastasis potential cell clones were more invasive and malignant than the low metastasis potential cell clones.