The Effect Of Breast Cancer Associated Fibroblast On Breast Caner Cells And Treatment Of Human Nasopharyngeal Carcinoma, Hepatocarcinoma And Thyroid Carcinoma Cell Subcutaneous Tumor Model By Oncolytic Herpes Simplex Virus Vector

ObjectiveTo study the effect of breast cancer associated fibroblast (CAFs) on breastcancer sells in vitro.MethodsThe CAFs and non-cancer associated fibrolasts (NAFs) were isolated fromprimary breast tumors and their paired normal breast tissue. The CAFs and NAFswere cultured in1640medium without the supplement of serum, and the supernatantwere collected.Breast cancer cell lines MCF-7and MDA-MB-468were cultured inthe collected supernatant, After48h of culture, the cells were collected andcalculated. Then the cell cycles were tested and the breast cancer stem cell markersCD44and CD24were tested by Flow Cytometry.ResultsWe successfully isolated and cultured the NAFs and CAFs. The CAFs increasedthe more expression ofα-smooth muscle actin (α-SMA) which is a definingcharacteristic of myofibroblasts. The proportion of α-SMA positive cells in NAFs and CAFs were6.00%±0.57%、53.33%±2.33%(P＜0.001) respectively. The CAFs andNAFs had no effect on breast cancer proliferation in vitro. After48h of culture, theproportion of G2/M phase of MCF-7were5.32%±0.02%、5.63%±0.03%、6.03±0.03%(P＞0.05)for the control group,NAFs group and CAFs grouprepectively. Both of the NAFs and CAFs increased the proportion ofCD44+CD24-cells in breast cancer cell line MCF-7and MDA-MB-468. the proportionof CD44+CD24-cells in MCF-7were0.41and2.51folds(P＜0.05) higher in the NAFsgroup and CAFs group. respectively than in the control group. And there were0.35and3.76folds(P＜0.05) higher in the MDA-MB-468respectively.ConclusionsThe CAFs highly express α-SMA, and had no effect on breast cancer proliferation invitro, but significantly increased the rates of cancer stem cells. The CAFs may play animportant role in the progresses of cancer recurrence and metastasis. ObjectiveTo investigate the efficacy of herpes simplex virus (HSV) vector, G47Δ for thetreatment of human nasopharyngeal carcinoma, hepatocarcinoma and thyroidcarcinoma through the observation of its treatment effect to subcutaneous tumor innude mice model. Methods and MethodsHuman nasopharyngeal carcinoma CNE-2, SUNE-1cells, hepatocarcinomaSMMC-7721, BEL7404cells and thyroid carcinoma ARO, FRO cells were culturedin vitro. Subcutaneous tumor model was established with Balb-c nude mice bysubcutaneous injection of human nasopharyngeal carcinoma cells, hepatocarcinomacells and thyroid carcinoma cells. When tumors were approximately5mm in maximaldiameter, mice were unilaterally(the left side) inoculated with50μl of2×107G47Δor virus buffer for control twice a week. Tumor size was measured by external caliperand tumor volume was calculated (V=a×b2/2). If the diameter of tumor reached18mm or when the animals appeared moribund as characterized by a rough hair coat,lethargy, hunched posture,or limited ambulatory movements in response tostimulation.the mouse were sacrificed. Compared the tumor size and the survival timeof the two groups.ResultsIn the subcutaneous tumor model of nasopharyngeal carcinoma and thyroidcarcinoma., comparing with the mock-treated group, the tumor in the virus-treatedgroup regressed completely. For the G47Δ-treated mice bearing sc SMMC-7721,BEL-7404tumors, the tumor of5and4mice regressed completely. In thesubcutaneous tumor model of nasopharyngeal carcinoma, hepatocarcinoma andthyroid carcinoma, The survival time of the virus-treated group was significantlylonger than mock-treated group.ConclusionsThe result suggest that intratumoral inoculation of G47Δ could effectively curesubcutaneous tumor model of nasopharyngeal carcinoma, hepatocarcinoma andthyroid carcinoma. This experiment offered clue for G47Δ’s application to clinicaltherapy in nasopharyngeal carcinoma, hepatocarcinoma and thyroid carcinoma.whichencourages further clinical trials.