Carcinoma-associated Fibroblasts Effects On Biological Behavior Of Colon Carcinoma LoVo Cells In Vivo

Objective To observe the effects of human colon carcinoma-associated fibroblasts(CAFs)on the biological behavior of LoVo colon cancer cells in vivo and investigate its role in colon cancer invasion and metastasis.Methods 1 The primary human colon CAFs and normal fibroblasts(NFs)were obtained by digestion methods.2 Identified the third generation CAFs and NFs cells by morphological observation and immunocytochemistry(ICC)staining.3 BALB / c nude mice were randomly grouped.Mix the above two cells with colon cancer LoVo cells at a concentration of 1×107cells / ml in a 1:1ratio and inject alone LoVo cells as the control group,and these cells are injected in spleen and subcutaneous to establish the subcutaneous xenotransplanted tumor model and liver metastasis model.Compare the differences among the growth rate of the three groups of subcutaneous tumors and liver metastasis rate.4 Detection of microvessel density(MVD)labeled CD31 by immunohistochemical SP method in the xenograft specimens to investigate CAFs,role in tumor angiogenesis.Results 1 ICC shows that FAP,FSP,Desmin are negative in NFs;Desmin is negative,FAP and FSP are positive in CAFs.2 Subcutaneous tumor growth curve shows the growth rate of CAFs + LoVo tumor volume is significantly greater than NFs + LoVo and LoVo group,and the difference was statistically significant(P<0.05).3 Immunohistochemistry shows the MVD count of CAFs + LoVo group are(11.16 ±2.44),MVD count of NFs + LoVo group are(7.36 ± 1.52),MVD count of LoVo group are(6.08 ± 1.41),MVD count of CAFs + LoVo group are significantly more than of the NFs +LoVo and LoVo group,and the difference is also statistically significant(P <0.05)between NFs + LoVo and LoVo group.4 The liver metastasis rate of CAFs + LoVo group is 80%,significantly higher than NFs + LoVo and LoVo group 40% to 20%,the difference was statistically significant(P <0.05).Conclusion 1 The purified human colon CAFs and NFs can be performed successfully by digestion methods.2 CAFs can promote human colon tumor angiogenesis.3 CAFs might promote human colon cancer invasion and metastasis.