In-vivo antiproliferative activity of Morus latifolia leaf and bark extracts against Ehrlich's ascites carcinoma

Abstract

Cancer is the second death causing disease all over the world and until today 100 different types of cancer have been identified whose treatment methods consist of serious side effects on human body. To reduce the frequency of adverse effects of cancer treatment, nowadays plant derived natural components are getting priority. The plant Morus latifolia is widely available in northern part of Bangladesh. The earlier researches suggested that popular varieties of some Morus sp. like Morus alba, Morus indica etc. have good anti-proliferative activity. Hence, this study was designed to evaluate the anti-proliferative activity of leaf and bark extracts of M. latifolia against Ehrlich's ascites carcinoma (EAC) in vivo. The leaf and bark extracts of M. latifolia were used in several bioassays including Brine shrimp lethality test, hemagglutination activity test, antioxidant activity test, and cell growth inhibition test. Besides, fluorescence microscopy was performed to study apoptotic features in EAC cells, and molecular analysis like real-time PCR were also conducted. The results of Brine shrimp lethality test, hemagglutination activity test, and antioxidant activity assay supported the cell growth inhibition capability of leaf and bark extracts which was confirmed by in vivo cell growth inhibition bioassay. Moreover, the experimental extracts were able to induce cell apoptotis through altering the expression pattern of Bcl-2 and Bax genes. All of the results of this study suggest that several noble compounds are present in M. latifolia plant extracts which are capable for healing cancer cells.

Keywords: Apoptosis; Bax; Bcl-2; Cancer; Cytotoxicity; Morus latifolia.

Fig. 1

Hemagglutination activity of leaf and bark extract of M. latifolia. The extracts agglutinate 2% RBC where control showed no agglutination. The leaf extract shows comparatively higher agglutination activity on mouse RBC indicating presence of more lectins than the bark extract

Fig. 2

Evaluation of cytotoxic activity of leaf and bark extract of M. latifolia. Mortality frequency of Brine shrimp nauplii treated by leaf and bark extract was increased in a dose-dependent manner. The data are represented as mean ± SD (n = 3)

Fig. 3

Antioxidant activity of leaf and bark extract of M. latifolia. Leaf extract of M. latifolia demonstrates the higher scavenging activity than bark extract when compare to standard (BHT). The results are represented as mean ± SD (n = 3)

Fig. 4

Observation of EAC cells for both control and treated mice under fluorescence microscope. The white arrows indicate the shrinkage, abnormal and irregular shaped in treated mice cells where round and regular shaped cells are appeared in control mice

Fig. 5

Average number of apoptotic cells per slide. Significant number of apoptotic cells was found in treated group compared to control group. The result is presented as mean ± SD (n = 3). Significance was set at *p < 0.05, **p < 0.01, and ***p < 0.001 in respect to control

Fig. 6

Effect of bark and leaf extract of M. latifolia on expression pattern of Bcl-2 and Bax gene in EAC cells in vivo. In case of bark, A = Bcl-2 and C = Bax. On the other hand, in case of leaf, B = Bcl-2 and D = Bax. The results are represented as mean ± SD (n = 3). Significance levels were determined and set at *p < 0.05, **p < 0.01, and ***p < 0.001 compared to the control