ABSTRACT
Objective To explore the impact of TIMP 3 regulated by miR-181b as a target gene on invasion and migration of hepatocellular carcinoma (HCC) in vitro.Methods The expressions of miR-181b were detected using SYBR Green real-time fluorescence quantitative polymerase chain reaction on liver cancer specimens and on HCC cell lines.The protein expression of TIMP 3 in HCC was detected using westen blot,and SKHep-1 as a cell line expressing high miR-181b was chosen through reporter gene experiment.TIMP-3 as a target gene regulated by miR-181b and its effect on invasion and migration treated by anti-miR-181 b were studied using transwell and cell scarification test,respectively.Results The expression of miR-181b in HCC was higher than cancer-adjacent tissues and normal liver tissues.The differences among them were significant.There was a correlation between the high expression of miR-181b and invasiveness and metastasis in HCC.The protein expression of TIMP-3 in HCC was significantly lower than normal liver tissues and cancer-adjacent tissues.Expression of miR-181b mRNA was detected in various HCC cell lines such as Hep3B,HepG2,Huh 7,SKHep-1,SNU182,SNU449 and hepatocyte,with the expression of miR-181b in SKHep-1 being the highest (P＜0.01).TIMP3-3UTR was low when the expression of miR-181b was high (P＜0.05).The invasion and migration abilities of SKHep-1 were significantly inhibited by anti-miR-181b (P＜0.05).Conclusion The data suggested that miR-181b promoted invasion and migration of SKHep-1 by down-regulating TIMP-3 in HCC.
ABSTRACT
<p><b>BACKGROUND</b>To investigate the outgrowth inhibition of the HPV16-positive murine lung tumor induced by a modified HPV16 mE6Δ/mE7 recombinant fusion protein vaccine in vivo and provide a new clue for the further immunotherapy.</p><p><b>METHODS</b>For prophylactic experiments, C57BL/6 mice were immunized with mE6Δ/mE7 fusion protein, and then inoculated with the TC-1 tumor cell, expressing HPV16 E6 and E7 viral proteins. On day 33 after inoculation, the tumor-free mice were re-challenged with a larger dose of TC-1 tumor cells. For therapeutic experiments, mice were vaccinated with mE6Δ/mE7 on days 3 and 14 after tumor cell inoculation. On day 60, the tumor-free mice were re-challenged with a larger dose of tumor cells. Tumor incidence and tumor volume of each group were calculated. MTT method was used to determine the proliferation of lymphocyte.</p><p><b>RESULTS</b>In the prophylactic experiments, immunization with the mE6Δ/mE7 completely protected the mice against the tumor cell challenge and rechallenge, and all the mice remained tumor free during the 100 days' observation period. In contrast, all the mice in PBS and IFA-treated groups developed tumors within 6-12 days after the first tumor cell inoculation, and died of tumor burden within 30 days. In the therapeutic experiments, the tumor formation rates were 20%, 90% and 60% in vaccinated, PBS and IFA groups respectively. In the next larger dose of tumor cells rechallenge experiment, 87.5% of vaccinated mice still remained tumor free, but all the mice from either PBS or IFA group developed tumors with 4-6 days. In addition, the results of MTT indicated that the proliferation of lymphocytes from vaccinated mice was stronger than that from control group.</p><p><b>CONCLUSIONS</b>The modified mE6Δ/mE7 can efficiently inhibit the growth of lung cancer in the animal model, indicating that mE6Δ/mE7 protein-based vaccine might show promise for the future clinical application.</p>
ABSTRACT
Purpose:To study the expression of uPA, uPAR ,PAI 1 in 3 nasopharyngeal carcinoma cell lines CEN 2Z,CEN 2Z H5,CEN 2Z H5 9,with different ability to metastasize through lymph.Methods:RT PCR was used to detect expression of uPA,uPAR,PAI 1 at levels of mRNA.Results:mRNA expression of uPA,uPAR was the highest in CNE 2Z H5 9,whereas the lowest in CNE 2Z;mRNA expression of PAI 1was detected in CNE 2Z and CNE 2Z H5, but revealed no differences,but not in CNE 2Z H5 9.Conclusions:uPA?uPAR may promote the metastasis and invasion of NPC,whereas PAI 1 maybe inhibit it