Experimental Study Of Inhibitory Effects Of Chloroquine On Hepatocellular Carcinoma And Its Mechanisms

Up to date, hepatocellular carcinoma therapy is still one of the serious problems that puzzle the scientists in the world. It is an important topic to seek an efficient way to treat hepatocellular carcinoma in the study of hepatocellular carcinoma. In this study, we explored the topic.1. Influence of chloroquine on apoptosis and cell cycle and the proliferation.First: HepG2 cells were divided into six groups:control group and chloroquine (8,16,32,64 and 128uM ) groups. The cell viability was determined by MTT; The cell cycle and apoptosis was detected by flow cytometry. Results showed that the cell viabilities were inhibited significantly 24h after chloroquine (32-128 uM) or 48-72h after chloroquine (8-128 uM) (P<0.01) ,compared with control group; HepG2 cells treated with chloroquine (32-128uM) for 24h were obviously arrested at G2/M phases,compared with control group (P<0.01). we concluded that chloroquine can suppress the activities of hepatoma HepG2 cells by inhibiting cells proliferation, inducing cells apoptosis and arresting cell cycle in vitro. So chloroquine is probably the drug for hepatocellular carcinoma.2. The study found that cell apoptosis was different with different chloroquine and blockers.The apoptosis increased when the cells were treated by chloroquine and PDTC;The apoptosis decreased when the cells were treated by chloroquine and sp600125;The apoptosis was not changed by chloroquine and SB203580 and PD98059.3. Measurement of protein levelFour protein were studied, including NF-кB,p-IкBa,P38,ERK and p-JNK. Western Blot assay was employed for the measurement of protein level 3.1 Changes in NF-кB protein expressionResults showed that NF-кB protein expression tended to decrease with the increase of chloroquine , the most effective inhibitory occurred at 128 uM. The inhibitor of NF-кB increased the chloroquine effect.3.2 Changes in p-IкBa protein expressionResults showed that p-IкBa protein expression tended to decrease with the increase of chloroquine , the most effective inhibitory occurred at 128 uM.3.3 Changes in P38 protein expressionResults showed that no any change was observed in P38 protein expression.3.4 Changes in p-JNK protein expressionResults showed that p-JNK protein expression tended to increase with the increase of chloroquine , the most effection occurred at 128 uM.3.5 Changes in ERK protein expression Results showed that ERK protein expression tended to decrease with the increase of chloroquine , the most effective inhibitory occurred at 128 uM. 3.6 By application of blockers, the study further confirmed that the apoptosis was increased due to the inhibitory of NF-кB and p-IкBαby chloroquine. The apoptosis was increased due to the increase of p-JNK by chloroquine.The proliferation of cells was inhibited owing to the ERK protein decrease by chloroquine.Taken together, the chloroquine can suppress the activities of hepatoma HepG2 cells by inhibiting cells proliferation, inducing cells apoptosis and arresting cell cycle in vitro. So chloroquine is probably the drug for hepatocellular carcinoma. By inhibiting the protein expression of NF-кB,p-JNK and ERK. But there was no change in the express of P38 protein.