Inhibition of tumor growth and metastasis with antisense oligonucleotides (Cantide) targeting hTERT in an in situ human hepatocellular carcinoma model

Abstract

Aim: To evaluate the in vivo antitumor effects of Cantide and the combined effect with 5‐fluorouracil. Methods: An in situ human hepatocellular carcinoma model was established in mice livers orthotopically. Drugs were administered intravenously and tumor sizes were monitored with calipers. Plasma alpha‐fetoprotein (AFP) were detected by radiation immunoassay. Morphology of tumors was evaluated by hematoxylin‐eosin (H&E) staining of histological sections. Human telomerase reverse transcriptase (hTERT) protein levels were detected by Western blotting. Results: Cantide significantly inhibit in situ human hepatocellular carcinoma growth in mice with a 75 and 50 mg·kg‐1·d‐1 administration of Cantide compared to the saline group in a dose‐dependent manner, which included injecting Cantide 25 mg·kg‐1·d‐1‐VS mg·kg‐1·d‐1 by iv for 20 d after surgically removing the tumor in liver. Cantide was also found to prevent tumor recurrence in the liver and metastasis in the lung, showing a dose‐dependent response. When Cantide was administered by iv combined with 5‐fluorouracil, it resulted in a significant reduction in tumor growth compared to either agent alone treatment group. After the treatment with Cantide alone or combined with 5‐fluorouracil, plasma AFP concentration decreased in a dose‐dependent manner. Conclusion: These results demonstrated that Cantide was an effective antitumor antisense oligonucleotide in vivo and has the potential to be developed into a clinical anti‐cancer drug.

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles

“…All animal experiments were carried out according to the standards of animal care as outlined in the NIH guide for the Care and Use of Laboratory Animals. The human HCC tumor model was described previously [17] . Briefly, the HCM-Y89 tumor derived from a surgical specimen of HCC was cut into 1 mm × 1 mm × 1 mm pieces, and implanted into the liver of mice.…”

“…The tumors were removed and fixed in neutral buffered 10% formalin, processed by standard methods, embedded in paraffin and sectioned and stained with HE. The HCC model maintains various important features similar to clinical liver cancer patients, including local growth, regional invasion, lymph nodes and pulmonary metastasis, peritoneal seeding with bloody ascites, and secretion of AFP in the recipient animals [17] .…”

Antisense oligonucleotides targeting midkine inhibit tumor growth in an in situ human hepatocellular carcinoma model

“…The tumors were removed and fixed in neutral buffered 10% formalin, processed by standard methods, embedded in paraffin, sectioned and stained with hematoxylin-eosin (HE). It is important to note that the HCC model maintains various features similar with clinical liver cancer patients including local growth, regional invasion, lymph nodes and pulmonary metastasis, peritoneal seeding with bloody ascites, and secretion of alpha-fetoprotein (AFP) in the recipient animals [29] . Treatment of in situ HCC xenograft with MK-AS and anticancer drugs Two days after the in situ HCC models were established, the tail veins of the mice were injected with saline (vehicle control), MK-AS (25, 50, and 100 mg·kg Histopathological analysis The tumor tissues were excised and fixed in 10% buffered formalin.…”

“…All animal experiments were carried out according to the standards of animal care as outlined in the NIH guide for the Care and Use of Laboratory Animals. The human HCC tumor model was described previously [29] . Briefly, the HCM-Y89 tumor derived from a surgical specimen of HCC was cut into 1 mm×1 mm×1 mm tissues, and implanted into the liver of mice.…”

“…Therefore, it is often used as an indicator of HCC [29] . In this experiment, we used RIA to detect serum AFP concentration at the endpoint of treatment.…”