In addition, TLR3 may directly trigger apoptosis in certain cancer cells. In addition, TLRs in tumor cells facilitate their evasion from immune surveillance via the suppression of T cell proliferation and natural killer cell activity, suggesting that TLR signaling www.selleckchem.com/products/VX-770.html in tumor cells is associated with the progression of cancer and evasion of host defenses. Thus, TLRs could be considered as potential therapeutic targets in HCC. Due to the complexity of genetic aberrations in HCC, single agent often fails to suppress tumor growth com pletely, therefore, combination of two or more anti cancer agents would greatly increase therapeutic effects. In this study, we evaluated the expression of a series of proteins relating to TLR3 signaling pathway, such as NFB, caspase 8 survivin, bcl2 and PCNA, affected by dsRNA or sorafenib alone or in combination of both.

According to the sequence of TLR3 sensitive virus, 4 dsRNAs were synthesizede, and only one was selected as a TLR3 synergist since it was the most effective to activate TLR3. Our results showed that combination of BM 06 with Sorafenib was superior to either agent alone in the inhibition of tumor growth either in vitro HCC cell lines or in vivo HCC rat models. Future investigations will be focused on the mechanism of the activated TLR3 in inhibiting HCC, and ultimately, a more effective anti HCC therapy will be evaluated using a combination use of sorafenib and dsRNA. Methods Cell culture Human HCC cell line producing HBV was purchased from Ruijin Hospital.

The cells were maintained in a Dulbeccos modified Eagles medium supplemented with 20% fetal bovine serum, 2 mM L glutamine, and 100 U mL penicillin streptomycin mixture at 37 C and 5% CO2 in a humidified chamber. Design and syntheses of 4 dsRNA TLR3 synergists Four dsRNAs were designed and synthesized by Bio mics. Co. Ltd. Their antisense se quences are as follows HepG2. 2. 15 cells were seeded into the wells of a 6 well culture plate and allowed to grow until 80% confluence. Subsequently, these cells were treated with the above 4 different dsRNAs, respectively. After treat ment at 37 C for 24 hours, the mRNA and protein ex pressions of TLR3 and NFB in HepG2. 2. 15 cells were measured by qRT PCR and Western blot, respectively. The dsRNA that showed the most effective activation against TLR3 interferon pathway was selected for fol lowing interventional treatments.

Treatments of HepG2. 2. 15 cell line HepG2. 2. 15 cells were incubated with the synthetic dsRNA, or sorafenib alone, or BM 06 plus sorafenib. In the present study, poly and phosphate buffered saline were used as a positive selleck screening library and a negative control, respect ively, by replacing dsRNA or sorafenib. Sorafenib obtained from Nantong Tomor Hospital in the commercially avail able form of 200 mg tablets was dissolved in 100% DMSO on the day of treatment.