Inhibition of Hepatocellular Carcinoma by Total Alkaloids of Rubus alceifolius Poir Involves Suppression of Hedgehog Signaling.
OBJECTIVE
We evaluated the effects of total alkaloids of Rubus alceifolius Poir (TARAP) on the migration and invasion of hepatocellular carcinoma (HCC) and furthermore investigated the possible molecular mechanisms mediating its anticancer activity.


METHODS
We implanted nude mice with human HCC HepG2 cells and fed them with vehicle (physiological saline) or 3 g/kg/day dose of TARAP 5 days per week for 21 days. We determined the in vitro effect of TARAP on the migration and invasion of HepG2 cells by transwell assay. We evaluated SHH signaling components' (SHH, PTCH, SMO, and Gli1) expression levels by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Activity of the matrix metalloproteinases (MMPs) in supernatants was analyzed by zymography. The expression of the MMPs and their specific tissue inhibitor (tissue inhibitor of matrix metalloproteinases, TIMP-1, 2) in HCC tissues was detected by immunohistochemistry.


RESULTS
We discovered that TARAP inhibited hepatocellular migration and invasion in a dose-dependent manner in vitro. In addition, TARAP decreased the expression of SHH, PTCH, SMO, and Gli1 in HCC mouse tumors at both transcriptional and translational levels. Moreover, TARAP inhibited the activity of MMP2 and MMP9. We found that TARAP reduced the expression of MMP2 and MMP9, as well as the tissue inhibitor of MMPs.


CONCLUSIONS
Our study showed that TARAP inhibits HCC migration and invasion likely through suppression of the hedgehog pathway. This may, in part, explain its anticancer properties. These results suggest that total alkaloids in Rubus alceifolius may have potential as a novel antimetastasis drug in the treatment of HCC.
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Jinyan
J
Zhao
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Liya
L
Liu
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Yun
Y
Wan
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Yuchen
Y
Zhang
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Qunchuan
Q
Zhuang
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Xiaoyong
X
Zhong
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China zfhong1953@163.com.
Zhenfeng
Z
Hong
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Jun
J
Peng
D000470:Alkaloids / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology; D002465:Cell Movement / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D053823:Hedgehog Proteins / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness / Q000517:prevention & control; D020133:Reverse Transcriptase Polymerase Chain Reaction; D065856:Rubus / Q000737:chemistry*; D015398:Signal Transduction / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
D000470:Alkaloids; D000972:Antineoplastic Agents, Phytogenic; D053823:Hedgehog Proteins
Hedgehog signaling pathway; hepatocellular carcinoma; invasion; migration; total alkaloids of Rubus alceifolius Poir