MiR-34a overexpression enhances the inhibitory effect of doxorubicin on HepG2 cells

BACKGROUND Hepatocellular carcinoma(HCC) is the third leading cause of death from malignant tumors worldwide. More than 50% of HCC cases occur in China. The prognosis remains poor and overall efficacy is still unsatisfactory. Chemotherapy resistance is the most important reason for the poor outcome. Much progress has been made in the study of chemotherapy resistance of HCC;however, the specific mechanisms of progression of HCC have still only been partially established.Therefore, the mechanism of chemotherapy resistance in HCC requires more research.AIM To investigate the effect of miR-34 a expression on the growth inhibition of HepG2 cells by doxorubicin.METHODS A recombinant lentiviral vector containing miR-34 a was constructed and transfected into HepG2 cells. The expression of miR-34 a was detected by reverse transcription-polymerase chain reaction(commonly known as RT-PCR) before and after transfection. Cells were exposed to 2 μM doxorubicin or phosphatebuffered saline before and after transfection. Cell viability in each group was detected by MTT assay, and cell cycle and apoptosis were detected by flow cytometry. Changes in expression levels of phospho(p)-p53, sirtuin(SIRT) 1,cyclin D1, cyclin-dependent kinase(CDK) 4, CDK6, BCL-2, multidrug resistance protein(MDR) 1/P glycoprotein(P-gp), and AXL were detected by Western blotting.RESULTS Recombinant lentiviral vector LV-hsa-mir-34 a was successfully constructed by restriction endonuclease digestion and sequencing. RT-PCR showed that expression of miR-34 a in HepG2 cells was significantly upregulated after transfection(P < 0.01). MTT assay showed that growth of HepG2 cells was inhibited after upregulation of miR-34 a, and viability was significantly decreased after combined treatment with doxorubicin(P < 0.01). Flow cytometry showed that the number of HepG2 cells in G1 phase increased, and G1 phase arrest was more obvious after intervention with doxorubicin(P < 0.01). The apoptosis rate of HepG2 cells was increased after upregulation of miR-34 a, and became more obvious after intervention with doxorubicin(P < 0.01). Western blotting showed that upregulation of miR-34 a combined with treatment with doxorubicin caused significant changes in the expression levels of p-p53, SIRT1, cyclin D1, CDK4,CDK6, BCL-2, MDR1/P-gp and AXL proteins(P < 0.01).CONCLUSION MiR-34 a may enhance the inhibitory effect of doxorubicin by downregulating MDR1/P-gp and AXL, which may be related to p53 expression.

Feasibility and safety of sorafenib treatment in hepatocellular carcinoma patients with spontaneous rupture

AIM:To report the outcome of patients with ruptured hepatocellular carcinoma(HCC)treated at a single center during a 5-year period.METHODS:We retrospectively analyzed 32 patients who presented with ruptured HCC at Shandong Provincial Hospital Affiliated to Shandong University between2008 and 2013.RESULTS:The mean age of the patients was 53 years(range 39-71 years).Of these patients,22 received surgical management,10 underwent transarterial embolization(TAE)or transarterial chemoembolization(TACE),and 12 received sorafenib after surgery,TAE or TACE.Cumulative survival rates at 4,8 and 12 mo were72.9%,50.0%and 33.3%,respectively,in the surgery only group and were 90.0%,80.6%and 64.1%,respectively,in the surgery plus sorafenib group.Cumulative survival rates at 4,8 and 12 mo were 68.4%,43.6%and 19.4%,respectively,in the surgery only or TAE/TACE only groups,and were 91.7%,75.0%and 60.2%,respectively,in the sorafenib combination groups(P=0.04).No unexpected side effects due to sorafenib were observed.The most common side effect was hand-foot skin reaction.To date,5 patients have died.Median follow-up from the start of sorafenib therapy for the remaining 7 patients is 12.7 mo(range5.8-32.2 mo).CONCLUSION:Sorafenib can be used in patients with ruptured HCC as it has interesting activity and is well tolerated;dose adjustment is generally not required.However,a larger prospective study is necessary to determine the efficacy of sorafenib in this group of patients.

Clinicopathological characteristics and surgical outcomes of sarcomatoid hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC)is the sixth most common type of cancer and the fourth leading cause of cancer-related death worldwide.Sarcomatoid HCC,which contains poorly differentiated carcinomatous and sarcomatous components,is a rare histological subtype of HCC that differs from conventional HCC.It is highly aggressive and has a poor prognosis.Its clinicopathological characteristics,surgical outcomes and underlying mechanisms of its highly aggressive nature have not been fully elucidated.AIM To examine the clinicopathological characteristics and surgical outcomes of sarcomatoid HCC and explore the histogenesis of sarcomatoid HCC.METHODS In total,196 patients[41 sarcomatoid HCC and 155 high-grade(Edmondson-Steiner grade III or IV)HCC]who underwent surgical resection between 2007 and 2017 were retrospectively reviewed.The characteristics and surgical outcomes of sarcomatoid HCC were compared with those of patients with high-grade HCC.The histological composition of invasive and metastatic sarcomatoid HCCs was evaluated.RESULTS Sarcomatoid HCC was more frequently diagnosed at an advanced stage with a larger tumor and higher rates of nonspecific symptom,adjacent organ invasion and lymph node metastasis than high-grade HCC(all P<0.05).Compared with high-grade HCC patients,sarcomatoid HCC patients are less likely to have typical dynamic imaging features of HCC(44.4%vs 72.7%,P=0.001)and elevated serum alpha-fetoprotein levels(>20 ng/mL;36.6%vs 78.7%,P<0.001).The sarcomatoid group had a significantly shorter median recurrence-free survival(5.6 mo vs 16.4 mo,log-rank P<0.0001)and overall survival(10.5 mo vs 48.1 mo,log-rank P<0.0001)than the high-grade group.After controlling for confounding factors,the sarcomatoid subtype was identified as an independent predictor of poor prognosis.Pathological analyses indicated that invasive and metastatic lesions were mainly composed of carcinomatous components.CONCLUSION Sarcomatoid HCC was associated with a more advanced stage,atypical dynamic imaging,lower serum alpha-fetoprotein levels and a worse prognosis.The highly aggressive nature of sarcomatoid HCC is perhaps mediated by carcinomatous components.