To Found Hela Mice Tumor Xenograft Model And The Effect Of Garlicin On Transpianted Tumor

Objective: Cervical carcinoma is a common malignant tumor. It's well known that HPV(Human papilloma virus) is the initiation factor in the occurrence of the cervical cancer. Up to now, HPV can't be cultivated in vitro, because of which, the research about preventing cervical cancer falls behind. The nude mice tumor xenograft model is one of the best method to study the character of the tumor and effect of the therapy. Based on previously study, the purpose of this study is to found Hela mice tumor xenograft model and study the effect of Garlicin on mice tumor xenograft, through which to provide a new way to prevent and cure the cervical cancer in clinic. Method 1 Hela cells frozen in liquid nitrogen were revived in routine methed and inoculated in RPMI-1640 blended with 10% foetus cattle serum. 2 4×10~6 human cervical cancer Hela cells were implanted into nude mice to get the tumor model of Hela cell line. The transplanted tumors were observed every two days. 3 Two weeks later after inoculating Hela cells, the mice were divided into four groups in random. 1.5ml Garlicin with different doses was vein-injected every other day, while 1.5ml saline solution was injected the control group mice every other day. After injecting 7 times the mice were killed the tumors were processed for several examination. 4 All groups mice blood was detected, as well as kidney and liver tissues were observed by optical microscope to find whether Garlicin had cytotoxic effect on the mice liver and kidney tissue. 5 After all groups transplanted tumors were fixuped dyhydrated embedded in olefin cut into pieces, the tumor pieces were stained with HE and observed by optical microscope to detect the morphologic change of the tumor cell sinduced by Garlicin. 6 All groups transplanted tumors were made section in routine. And the slices were observed by electron microscope to detect the apoptosis induced by Garlicin. 7 All groups transplanted tumors slices were treated according to instruction of TUNEL test kit. The cells whose nucleuses were stained with brown were apoptosis cells, while the blue ones were negative. 100 cells were counted in every section and then calculated apoptosis rate of cells. 8 With PCR, the expression change of HPV18-E6 DNA which had combined with DNA of Hela cell was studied in vivo induced by Garlicin. The HPV18-E6 level was expressed with HPV18-E6/β-actin. Result 1 Three days after transplanting Hela, all mice becamelazy and thin, and 15 mices grow transplanted tumor. And seven days later, all groups transplanted tumors were available. 2 Comparing with the control group, when the mass concentrations of Garlicin were 10mg/kg,25mg/kg,40mgk/g the tumor inhibiting rates were 33.32%,53.58%,82.46% (P﹤0.05)( Tabel 1). 3 Comparing with the control group, the ALT,AST,BUN and Cre in the blood serum hadn't remarkable difference in all Garlicin groups. Garlicin didn't affect the liver and kidney function. And morphologic kidney and liver tissues had no chang(Tabel 2,Tabel 3). 4 In control groups, Hela cells had few apoptosis cells, and organelle was integrated. In trial groups, smaller cells concentrated cytoplasy and expanding rough endoplasmic reticulum were seen. Chromatin assemblaged to blocks on the karyotheca(Fig.9). 5 The apoptosis rates of the control group and three different concentration groups were 3.93±1.19%,6.40±0.80%,9.02±0.93%,12.07±0.98%(P﹤0.05)(Tabel 4). 6 In trial and control groups DNA, the HPV18-E6 was all expressed. The result showed that Garlicin can down-regulate the expression of HPV18-E6 in Hela cells in vivo(P ﹤0.05)( Fig.12,Tabel 5). Conclusion: Transplanted tumor of Hela may be a effective model to study the character of tumor and effection of therapy method. The result suggested that Garlicin could inhibit thetumor proliferation and induce to apoptosis. And it also can make decreased expression of HPV18-E6 in Hela cell.