Inhibition Of NDV7793on The Hepatic Metastases Of Colonic Adenocarcinoma In The Mice

Colorectal cancer was the third most commonly diagnosed cancer in malesand the second in females, with over1.2million new cancer cases and608700deaths occurred in2008. About60%-70%those patients died with hepaticmetastasis of colon cancer. Most part of the patients who could not meet thesurgery condition, therefore adequated for chemotherapy. But the toxic and sideeffects of chemotherapy are big problem as we know. So, the doctors put theireyes on the biotherapy. Targeted therapy is one of the tumor biotherapies whichcan reduce untoward effects of the chemotherapy. Though many targetedmedicines like Cetuximab and panitumumab recommend by NCCN Guidelines,researches and developments of new targeted medicines are still underway.Viral therapy is one of the biological anti-tumor therapies of tumor, theanti-tumor viruses has been clinically evaluated including adenovirus, herpessimplex virus, reovirus, vaccinia virus, and Newcastle disease virus. Newcastledisease virus (NDV) belongs to Avulavirus genera of the Paramyxoviridaefamily. Since anticancer potential of NDV was fond, the study of NDV antitumor has become an enduring topic. The specific killing effect of NDVontumor cells is verified by basic studies and clinic trials. The antitumormechanisms of NDV become gradually clear and systematic. Clinical test showsthat the autologous tumor modified by NDV could restrain the hepaticmetastases of colorectal cancer. NDV has become a candidate for NCI antitumordrugs in USA. Our team involves in the antitumor basic research with NDV7793since2004. We proved that NDV7793could kill the human colorectal cancerboth in vitro and vivo.5-fluorouracil has been used on clinical treatment ofcolorectal cancer for50years as a basic chemotherapy drug. Recently, anincreasing number of colorectal cancer patients who are esistant to5-fluorouracil leading to reduction of the5-fluorouracil curative effect. As aconsequence, the studies in all kinds of drugs combined with5-fluorouraciltherapy on colorectal cancer are springing up, and yielding gratifying fruits.This task is to study if NDV7793could inhibit liver metastases ofcolorectal cancer, and whether the combined effect exists with NDV7793and5-Fu. Meanwhile, we study the effect on immunity of the tumor-beard micecaused by NDV7793. And all we want to do is to offer firm evidences to theclinic use of NDV7793.Methods1. Establishment of colon cancer liver metastases mice moldel.Conventional cultured mice colon cancer CT26cell. To establish the livermetastasis model, we used about1×106CT26cells for intrasplenic injection.After the naturally death of mice model, the status of liver metastases wererecorded. HE staining method was used to stain the livers for pathologicalobservation. 2. The inhibition of NDV7793on murine colon carcinoma liver metastasis.NDV7793was inoculated in SPF level chick embryo for virusamplification, and was purified by ultracentrifugation. The liver metastasesmodels randomly allocate to four groups: PBS negative control group,5-Fupositive control group, NDV7793group and combination group. There were20mice in each group. All of them were given medicines by intraperitonealinjection (0.1ml) once a day. The process began in the day when the modelswere built and lasted for10days. Used repeated measurement method tocompare the changes of mice weight in different groups, and then analyzed thesurvival time of the tumor-bearing mice in each group.The remainder was killedin the6thday after drug withdrawal. Weighed and visual studied the livers of thekilled mice to contrast the liver metastasis level of colorectal cancer withdifferent group. Kept the whole livers for routine embedding, sectioning, HEstaining. Then analyzed pathological changes of the liver tissues under lightmicroscope, and counted the neoplastic foci in livers.3. The influence of NDV7793on the liver metastasis mice immunity.Established colon cancer liver metastasis models and grouped formedicines as above mentioned. There were10mice in each group. All of themice were executed in the6thday after the drug discontinuance. The mice andtheir thymuses were weighted to compare the thymus index of each group. Setup a normal group that randomizing acquired10health mice of the same age,given intraperitoneal injection of0.1ml PBS every day for10days, and killthem all in the6thday after the drug discontinuance. Glass slurry method wasused to obtain the liver tissue homogenate. IL-2, IFNγ, TNFα, IL-17in the livertissue homogenate of different groups was tested by ELISA. Results1. Successfully established the colorectal cancer hepatic metastasis model, thesuccessful rate reached90%(27/30).2. The inhibition of NDV7793on murine colon carcinoma liver metastases.1) Compared to PBS negative control group, NDV7793postponed theincrease of mice weight. The mice weight was losing while5-Fu wasgiven in5-Fu positive control group. In the combination group, the miceweight grew slow down when NDV7793was given, and then declinedwhile5-Fu was given.2) The survival time of NDV7793group was shorter than combination group,and shorter than that of combination group (P=0.000<0.001), and longerthan that of5-Fu positive control group and PBS negative control group(P=0.000<0.001).3) The anti-tumor rates revealed that the liver metastasis of colorectal cancerwere suppressed within16days after the tumor models were built inNDV7793group, combination group, and5-Fu positive control group.The anti-tumor rates of NDV7793group and the combination group wereall higher than that of5-Fu positive control group (P=0.000<0.01). Therewere no difference between NDV7793group anti-tumor rates andcombination group anti-tumor rates (P=0.871>0.05).4) The calculation of neoplastic foci in livers of NDV7793group were lessthan that of PBS negative control group and5-Fu positive control group(P=0.000<0.01). There were no difference between NDV7793groupneoplastic foci calculation and combination group neoplastic foci calculation (P=0.733>0.05).5) Visual study of the livers showed that almost the whole livers werecovered with neoplasm in PBS negative control group. And the scope ofcancer in5-Fu positive control group livers reached90%. But inNDV7793group and combination group, the neoplasm scopes thatcovered on the liver surface were only about50%.6) Pathological examination showed that that were more and biggerapoptosis/necrosis regions in neoplastic foci of NDV7793group andcombination group than PBS negative control group.3. The influence of NDV7793on the liver metastasis mice immunity.1) The thymus index of NDV7793group is higher than that of combinationgroup (P=0.004<0.01). The thymus index of combination group is higherthan that of PBS negative control group and5-Fu positive control group(P=0.000<0.01).2) The expression of IL-2in mice livers of NDV7793group is higher thanthat of PBS negative control group and5-Fu positive control group(P=0.000<0.05), and is lower than that of combination group(P=0.000<0.05).3) IFNγ tested in mice liver tissues showed that the livers of NDV7793group expressed more IFNγ than PBS negative controlgroup(P=0.000<0.05) and5-Fu positive control group(P＝0.005＜0.05).The IFNγ level of NDV7793group was lower than combination group(P=0.000<0.05).4) TNFα tested result showed that the levels of TNFα in the liver tissues ofNDV7793group was higher than that of PBS negative control group (P=0.000<0.05) and5-Fu positive control group (P=0.000<0.05), and waslower than combination group (P=0.000<0.05).5) ELISA showed that the IL-17in NDV7793group was higher than PBSnegative control group (P=0.000<0.05), and was lower than combinationgroup (P=0.000<0.05), but was no different with5-Fu positive controlgroup (P=0.074>0.05).Conclusions1. Successfully established the colorectal cancer hepatic metastasis model.2. The single used of NDV7793was better than5-Fu on the inhibition of micecolorectal cancer liver metastasis. With the combination of5-Fu, survival oftumor-bear mice has been extended, but antitumor effect had not been enhanced.3. NDV7793could improve the tumor-burdened mice immunity. Theup-regulation of NDV7793on the levels of IL-2, IFNγ, TNFα, IL-17in the liverof colon cancer liver metastases model was better when combined with5-Fu.