**6. Acknowledgment**

90 Hepatocellular Carcinoma – Basic Research

cytoplasmic vesicles-like and lysosome-like structures, suggesting that processing of the HA-drug molecules might be coupled with a recycling of the CD44 receptor (**Fig. 7B**). We have also demonstrated that, using this drug delivery strategy, the delivered drug exerts a strong and irreversible *in vitro* inhibitory effect on growth of CD44 over-expressing cancer

As mentioned above, the effectiveness of the drug delivery strategy using HA as a carrier targeting its CD44 receptor has been demonstrated in several kinds of CD44 over-expressing tumors, including colorectal, ovarian, bladder, gastric, breast, oesophageal and lung cancers, not only *in vitro* but also *in vivo* (Banzato et al., 2008; Luo & Prestwich, 1999; Luo et al., 2000; Luo et al., 2002; Rosato et al., 2006; Serafino et al., 2011). Due the association of increased expression of CD44 to advanced stages of hepatocellular carcinoma, this therapeutic approach might be also applicable for treatment of metastatic HCC. In addition, the growing evidences concerning the CD44 expression on liver cancer stem cells (Liu et al., 2011) not only improve the prognostic significance of CD44 but also make the drug delivery strategy through CD44/HA binding interaction useful for eradicating hepatic cancer stem

Hepatocellular carcinoma is a malignancy having multifactorial etiology and a very complex pathogenesis, that make difficult the clinical management of HCC patients. Similarly to the other kinds of cancer, HCC insurgence, progression and recurrence involve gene mutations, that might be different depending on the individual genotype profiling and tumor stage, and different signaling pathways, which often share some crucial molecules/steps and are subjected to additional post-transductional regulation. To overcome the complex network of signaling pathways and gene mutations underlying hepatic cancer, innovative diagnostic, prognostic and therapeutic strategies are needed. Nowadays, each existing biomarker used or proposed for HCC early diagnosis, staging and prognosis alone is poorly specific and the absolute positive and negative serological and/or immunohistochemical markers are still lacking. Even those markers selected for high sensitivity and specificity do not exhibit an universal diagnostic/prognostic value, also due to the individual genotype variability. The more promising approach for developing more specific diagnostic/prognostic tools might be to combine several positive or negative indicators in multiparametric platforms, that allow simultaneous detection of multiple serological or immunohistochemical markers for HCC. These platforms might be used to design "specific maps" for HCC early diagnosis, staging and prognosis, also taking into account the individual variability of each patient. Detecting expression patterns of combined biomarkers may also be a new method useful for

Moreover, since target-specific cancer therapy has remarkably improved the outcomes of patients and represents the frontline approach for cancer treatment, the development of such multiparametric platforms would also represent a high-performance technological tools useful for designing personalized therapies, adapted to the aggressiveness of each individual tumor. The final goal should be to discriminate, for each target-specific therapy and on the basis of the "biomarker profiling" of each patient, the "responder" to the "not responder", thus increasing the therapeutic effectiveness, improving patient outcomes, and resulting in saving healthcare costs. Thus, the discovery and validation of new HCC

cells, higher than that exerted by free drug.

cells.

**5. Conclusion** 

identifying new and unique markers.

We would like to thank Federica Andreola, Noemi Moroni and Manuela Zonfrillo, for their valuable contribution to the researches carried out in our laboratory on NGF and on CD44 and to the ongoing study on the role of WNT/β-catenin signaling in cancer. We are also grateful to Dr. Giuseppe Nicotera for his scientific secretariat support.
